U.S. patent number 8,580,798 [Application Number 12/158,524] was granted by the patent office on 2013-11-12 for substituted pyrimidine derivatives useful in the treatment of cancer and other disorders.
This patent grant is currently assigned to Bayer Intellectual Property GmbH. The grantee listed for this patent is Jacques Dumas, Wendy Lee, Karl Miranda, Roger Smith, Gan Wang. Invention is credited to Jacques Dumas, Wendy Lee, Karl Miranda, Roger Smith, Gan Wang.
United States Patent |
8,580,798 |
Smith , et al. |
November 12, 2013 |
**Please see images for:
( Certificate of Correction ) ** |
Substituted pyrimidine derivatives useful in the treatment of
cancer and other disorders
Abstract
Substituted pyrimidine derivatives of formula (I), salts,
metabolites, prodrugs and diastereoisomeric forms (both isolated
stereoisomers and mixtures of stereoisomers) thereof (wherein
A=pyrimidine) pharmaceutical compositions containing such compounds
and the use of those compounds or compositions for treating
hyper-proliferative and angiogenesis disorders, as a sole agent or
in combination with other active ingredients, e.g., cytotoxic
therapies. ##STR00001##
Inventors: |
Smith; Roger (Chester Springs,
PA), Dumas; Jacques (Carlisle, MA), Wang; Gan
(Wallingford, CT), Lee; Wendy (San Ramon, CA), Miranda;
Karl (Lexington, MA) |
Applicant: |
Name |
City |
State |
Country |
Type |
Smith; Roger
Dumas; Jacques
Wang; Gan
Lee; Wendy
Miranda; Karl |
Chester Springs
Carlisle
Wallingford
San Ramon
Lexington |
PA
MA
CT
CA
MA |
US
US
US
US
US |
|
|
Assignee: |
Bayer Intellectual Property
GmbH (Monheim, DE)
|
Family
ID: |
38091685 |
Appl.
No.: |
12/158,524 |
Filed: |
December 20, 2006 |
PCT
Filed: |
December 20, 2006 |
PCT No.: |
PCT/US2006/048382 |
371(c)(1),(2),(4) Date: |
September 22, 2009 |
PCT
Pub. No.: |
WO2007/075650 |
PCT
Pub. Date: |
July 05, 2007 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20100081812 A1 |
Apr 1, 2010 |
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Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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60752200 |
Dec 21, 2005 |
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Current U.S.
Class: |
514/256; 544/296;
544/328 |
Current CPC
Class: |
C07D
401/12 (20130101); A61P 35/00 (20180101); A61P
43/00 (20180101) |
Current International
Class: |
C07D
401/12 (20060101); A61K 31/506 (20060101) |
Field of
Search: |
;544/296,328
;514/256 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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1 777 218 |
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Apr 2007 |
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EP |
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2003-526613 |
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Sep 2003 |
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JP |
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2005-272474 |
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Oct 2005 |
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JP |
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WO 99/32106 |
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Jul 1999 |
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WO |
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WO 99/32110 |
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Jul 1999 |
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WO |
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WO 99/32111 |
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Jul 1999 |
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WO |
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WO 99/32455 |
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Jul 1999 |
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WO |
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WO 00/41698 |
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Jul 2000 |
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WO |
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WO 00/42012 |
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Jul 2000 |
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WO |
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WO 02/062763 |
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Aug 2002 |
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WO |
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WO 02/085857 |
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Oct 2002 |
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WO |
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WO 2004/078128 |
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Sep 2004 |
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WO |
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WO 2004/078746 |
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Sep 2004 |
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WO |
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WO 2004/078747 |
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Sep 2004 |
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WO |
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WO 2005/075425 |
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Aug 2005 |
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WO |
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WO 2006/071940 |
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Jul 2006 |
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WO |
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|
Primary Examiner: Rao; Deepak
Attorney, Agent or Firm: Millen, White, Zelano &
Branigan, P.C.
Parent Case Text
This application claims the benefit of U.S. Provisional Application
No. 60/752,200, filed Dec. 21, 2005.
Claims
What is claimed is:
1. A compound of formula (I), a salt thereof, an oxidized
derivative thereof, wherein one or more of the nitrogens are
substituted with a hydroxy group, or a diastereoisomeric form
thereof, either as an isolated stereoisomer or in a mixture of
stereoisomers ##STR00026## wherein A is pyrimidine, optionally
substituted with 1 to 3 substituents which are independently
R.sup.1, OR.sup.1, S(O).sub.pR.sup.1, C(O)R.sup.1, C(O)OR.sup.1,
C(O)NR.sup.1R.sup.2, halogen, hydroxy, amino, cyano, or nitro; B is
phenyl, naphthyl, or pyridyl, optionally substituted with 1 to 4
substituents which are independently C.sub.1-C.sub.5 linear or
branched alkyl, C.sub.1-C.sub.5 linear or branched haloalkyl,
C.sub.1-C.sub.3 alkoxy, hydroxy, amino, C.sub.1-C.sub.3 alkylamino,
C.sub.1-C.sub.6 dialkylamino, halogen, cyano, or nitro; L is a
bridging group which is: (a)
--(CH.sub.2).sub.m--O--(CH.sub.2).sub.l--, (b)
--(CH.sub.2).sub.m--(CH.sub.2).sub.l--, (c)
--(CH.sub.2).sub.m--C(O)--(CH.sub.2).sub.l--, (d)
--(CH.sub.2).sub.m--NR.sup.3--(CH.sub.2).sub.l--, (e)
--(CH.sub.2).sub.m--NR.sup.3C(O)--(CH.sub.2).sub.l--, (f)
--(CH.sub.2).sub.m--S--(CH.sub.2).sub.l--, or (g)
--(CH.sub.2).sub.mC(O)NR.sup.3--(CH.sub.2).sub.l--, where the
integers m and l are independently selected from 0-4 and for
--(CH.sub.2).sub.m--(CH.sub.2).sub.l-- m and l cannot both be 0; M
is a pyridine or pyrimidine ring, optionally substituted with 1-3
substituents which are independently selected from: (1)
C.sub.1-C.sub.5 linear or branched alkyl; (2) C.sub.1-C.sub.5
linear or branched haloalkyl; (3) C.sub.1-C.sub.3 alkoxy; (4)
hydroxy; (5) amino; (6) C.sub.1-C.sub.3 alkylamino; (7)
C.sub.1-C.sub.6 dialkylamino; (8) halogen; (9) nitro; (10) C(O)
NR.sup.4R.sup.5; (11) C(O)OR.sup.4; (12) C(O)R.sup.4; (13) CN; (14)
C(S)NR.sup.4R.sup.5; (15a) C(O)NR.sup.7--NR.sup.4R.sup.5; (15b)
C(O)NR.sup.7--R.sup.4--C(O)NR.sup.4R.sup.5; (16) tetrazolyl; (17)
imidazolyl; (18) imidazoline-2-yl; (19) 1,3,4-oxadiazoline-2-yl;
(20) 1,3-thiazoline-2-yl; (21)
5-thioxo-4,5-dihydro-1,3,4-thiazoline-2-yl; (22)
5-oxo-4,5-dihydro-1,3,4-oxadiazoline-2-yl; or (23) a group of the
formula ##STR00027## each of R.sup.1, R.sup.2, R.sup.3, R.sup.4 and
R.sup.5 is independently (a) hydrogen, (b) C.sub.1-C.sub.5 linear,
branched, or cyclic alkyl, (c) up to per-halo substituted
C.sub.1-C.sub.5 linear or branched alkyl, or (d)
--(CH.sub.2).sub.q--X. where the substituent X is a 5 or 6 membered
heterocyclic ring, containing at least one atom selected from
oxygen, nitrogen and sulfur, which is saturated, partially
saturated, or aromatic, or a 8-10 membered bicyclic heteroaryl
having 1-4 heteroatoms selected from the group consisting of O, N
and S; R.sup.2, R.sup.3, R.sup.4 and R.sup.5 may, independently,
additionally include phenyl or C.sub.1-C.sub.3 phenyl-alkyl;
R.sup.4 and R.sup.5 may optionally be taken together to form a 5 or
6 membered aliphatic ring, which may be interrupted by an atom
selected from N, O or S which is optionally substituted with 1-3
substituents which are independently C.sub.1-C.sub.5 linear or
branched alkyl, up to perhalo substituted C.sub.1-C.sub.5 linear or
branched alkyl, C.sub.1-C.sub.3 alkoxy, hydroxy, oxo, carboxy,
amino, C.sub.1-C.sub.3 alkylamino, C.sub.1-C.sub.6 dialkylamino,
halogen, cyano, or nitro; R.sup.6 is independently: (a) hydrogen,
(b) C.sub.1-C.sub.5 linear, branched, or cyclic alkyl, (c) cyano,
(d) nitro, (e) up to per-halo substituted C.sub.1-C.sub.5 linear or
branched alkyl, or (f) --C(O)R.sup.7, where R.sup.7 is
C.sub.1-C.sub.5 linear, branched, or cyclic alkyl; R.sup.7 is
hydrogen, or C.sub.1-C.sub.5 linear, branched, or cyclic alkyl; the
variable q is an integer 1, 2, 3, or 4 and the variable p is an
integer 0, 1, or 2.
2. A compound of claim 1 wherein B is phenyl, optionally
substituted with 1-4 substituents which are independently
C.sub.1-C.sub.5 linear or branched alkyl, C .sub.1-C.sub.5 linear
or branched haloalkyl, C.sub.1-C.sub.3 alkoxy, hydroxy, amino,
C.sub.1-C.sub.3 alkylamino, C.sub.1-C.sub.6 dialkylamino, halogen,
cyano, or nitro.
3. A compound of claim 1 or 2 where L is --O-- or --S--.
4. A compound of claim 1 , where M is pyridine, optionally
substituted with 1-3 substituents which are independently selected
from the groups (1) C.sub.1-C.sub.5 linear or branched alkyl; (2)
C.sub.1-C.sub.5 linear or branched haloalkyl; (3) C.sub.1-C.sub.3
alkoxy; (4) hydroxy; (5) amino; (6) C.sub.1-C.sub.3 alkylamino; (7)
C.sub.1-C.sub.6 dialkylamino; (8) halogen; (9) nitro; (10) C(O)
NR.sup.4R.sup.5; (11) C(O)O R.sup.4; (12) C(O) R.sup.4; (13) CN,
(15a) C(O)NR.sup.7--NR.sup.4R.sup.5; or (15b)
C(O)NR.sup.7--R.sup.4--C(O)NR.sup.4R.sup.5.
5. A compound of claim 1 where R.sup.6 is independently: (a)
hydrogen, (b) C.sub.1-C.sub.5 linear, branched, or cyclic alkyl, or
(c) cyano or (d) nitro.
6. A compound of claim 1 where R.sup.6 is independently: (a)
hydrogen, (b) C.sub.1-C.sub.5 linear, branched, or cyclic alkyl, or
(c) cyano.
7. A compound of claim 1 of formula (III), a salt thereof, an
oxidized derivative thereof, wherein one or more of the nitrogens
are substituted with a hydroxy group, or a diastereoisomeric form
thereof, either as an isolated stereoisomer or in a mixture of
stereoisomers, ##STR00028## wherein Ra is R.sup.1, OR.sup.1 or
cyano; and B, L and M are as defined in claim 1.
8. A compound of claim 1 of formula (IV), a salt thereof, an
oxidized derivative thereof, wherein one or more of the nitrogens
are substituted with a hydroxy group, or a diastereoisomeric form
thereof, either as an isolated stereoisomer or in a mixture of
stereoisomers, ##STR00029## wherein Ra is R.sup.1, OR.sup.1 or
cyano; each Rc is independently hydrogen, halogen, C.sub.1-C.sub.3
linear or branched alkyl, C.sub.1-C.sub.3 linear or branched
haloalkyl, C.sub.1-C.sub.3 alkoxy or hydroxy; and L and M are as
defined in claim 1.
9. A compound of claim 1 of formula (V), a salt thereof, an
oxidized derivative thereof, wherein one or more of the nitrogens
are substituted with a hydroxy group, or a diastereoisomeric form
thereof, either as an isolated stereoisomer or in a mixture of
stereoisomers, ##STR00030## wherein Ra is R.sup.1, OR.sup.1 or
cyano; each Rc is independently hydrogen, halogen, C.sub.1-C.sub.3
linear or branched alkyl, C.sub.1-C.sub.3 linear or branched
haloalkyl, C.sub.1-C.sub.3 alkoxy or hydroxy; and M is as defined
in claim 1.
10. A compound as in claim 8 or 9 wherein M is pyridine.
11. A compound as in claim 8 or 9 where in M is pyridine
substituted by C(O) NR.sup.4R.sup.5 or CN.
12. A compound as in claim 8 or 9 where in M is pyridine
substituted by C(O) NR.sup.4R.sup.5.
13. A compound as in claim 8 or 9 where in M is pyridine
substituted by C(O) NHCH.sub.3 or C(O)NH.sub.2.
14. A compound of formula (II), a salt thereof, an oxidized
derivative thereof, wherein one or more of the nitrogens are
substituted with a hydroxy group, or a diastereoisomeric form
thereof, either as an isolated stereoisomer or mixture of
stereoisomers, ##STR00031## wherein Ra is R.sup.1, OR.sup.1 or
cyano each Rc is independently hydrogen, halogen, C.sub.1-C.sub.3
linear or branched alkyl, C.sub.1-C.sub.3 linear or branched
haloalkyl, C.sub.1-C.sub.3 alkoxy or hydroxy; Rb is (1)
C.sub.1-C.sub.5 linear or branched alkyl; (2) C.sub.1-C.sub.5
linear or branched haloalkyl; (3) C.sub.1-C.sub.3 alkoxy; (4)
hydroxy; (5) amino; (6) C.sub.1-C.sub.3 alkylamino; (7)
C.sub.1-C.sub.6 dialkylamino; (8) halogen; (9) nitro; (10) C(O)
NR.sup.4R.sup.5; (11) C(O)OR4; (12) C(O)R.sup.4; (13) CN; (14)
C(S)NR.sup.4R.sup.5; (15) C(O)NR.sup.7--R.sup.4C(O)N
R.sup.4R.sup.5; or (16) hydrogen; R.sup.1 is hydrogen or
C.sub.1-C.sub.5 linear, branched, or cyclic alkyl, each of R.sup.4
and R.sup.5 is independently (a) hydrogen, (b) C.sub.1-C.sub.5
linear, branched, or cyclic alkyl or (c) phenyl, and R.sup.7 is
hydrogen, or C.sub.1-C.sub.5 linear, branched, or cyclic alkyl.
15. A compound of claim 14 wherein Rb is C.sub.1-C.sub.5 linear or
branched alkyl; C.sub.1-C.sub.3 alkoxy; halogen; C(O)
NR.sup.4R.sup.5; CN; C(S)NR.sup.4R.sup.5 or
C(O)NR.sup.7--R.sup.4C(O)N R.sup.4R.sup.5.
16. A compound of claim 14 wherein Rb is C.sub.1-C.sub.5 linear or
branched alkyl; halogen; C(O) NR.sup.4R.sup.5 or CN.
17. A compound of claim 14 , 15 or 16 wherein Rc is independently
hydrogen or fluorine.
18. A compound of claim 14 wherein Rb is C(O) NR.sup.4R.sup.5 or
CN.
19. A compound of claim 14 wherein Rb is C(O) NR.sup.4R.sup.5.
20. A compound of claim 14 wherein Rb is C(O) NHCH.sub.3 or C(O)
NH.sub.2.
21. A compound of claim 1 which is:
4-{3-fluoro-4-[3-(6-trifluoromethylpyrimidin-4-yl)ureido]phenoxy}pyridine-
-2-carboxylic acid methylamide,
4-{4-[3-(6-trifluoromethyl-pyrimidin-4-yl)-ureido]-phenoxy}-pyridine-2-ca-
rboxylic acid methylamide,
4-{4-[3-(6-trifluoromethylpyrimidin-4-yl)ureido]phenoxy}-2-methylpyridine-
,
1-[2-Fluoro-4-(2-methylpyridin-4-yloxy)phenyl]-3-(6-trifluoromethylpyrim-
id-in-4-yl)urea,
1-(6-tert-Butylpyrimidin-4-yl)-3-[4-(2-cyanopyridin-4-yloxy)-2-fluorophen-
y-l]urea,
4-{3-fluoro-4-[3-(6-tert-butylpyrimidin-4-yl)ureido]phenoxy}pyri-
dine-2-carboxylic acid methylamide,
4-{4-[3-(6-tert-Butylpyrimidin-4-yl)ureido]-3-fluorophenoxy}pyridine-2-ca-
rboxylic acid amide,
4-{4-[3-(6-tert-Butylpyrimidin-4-yl)ureido]phenoxy}pyridine-2-carbothioic
acid amide,
4-{3-fluoro-4-[3-(6-methoxypyrimidin-4-yl)ureido]phenoxy}pyridine-2-carbo-
xylic acid methylamide, or
4-{4-[3-(6-Phenylpyrimidin-4-yl)ureido]phenoxy}pyridine-2-carbothioic
acid amide.
Description
FIELD OF THE INVENTION
This invention relates to novel compounds, pharmaceutical
compositions containing such compounds and the use of those
compounds or compositions for treating hyper-proliferative and
angiogenesis disorders, as a sole agent or in combination with
other active ingredients, e.g., cytotoxic therapies.
BACKGROUND OF THE INVENTION
Activation of the ras signal transduction pathway indicates a
cascade of events that have a profound impact on cellular
proliferation, differentiation, and transformation. Raf kinase, a
downstream effector of ras, is recognized as a key mediator of
these signals from cell surface receptors to the cell nucleus
(Lowy, D. R.; Willumsen, B. M. Ann. Rev. Biochem. 1993, 62, 851;
Bos, J. L. Cancer Res. 1989, 49, 4682). It has been shown that
inhibiting the effect of active ras by inhibiting the raf kinase
signaling pathway by administration of deactivating antibodies to
raf kinase or by co-expression of dominant negative raf kinase or
dominant negative MEK, the substrate of raf kinase, leads to the
reversion of transformed cells to the normal growth phenotype (see:
Daum et al. Trends Biochem. Sci. 1994, 19, 474-80; Fridman et al.
J. Biol. Chem. 1994, 269, 30105-8. Kolch et al. (Nature 1991, 349,
426-28) have further indicated that inhibition of raf expression by
antisense RNA blocks cell proliferation in membrane-associated
oncogenes. Similarly, inhibition of raf kinase (by antisense
oligodeoxynucleotides) has been correlated in vitro and in vivo
with inhibition of the growth of a variety of human tumor types
(Monia et al., Nat. Med. 1996, 2, 668-75). Some examples of small
molecule inhibitors of Raf kinase activity are important agents for
the treatment of cancer. (Naumann, U.; Eisenmann-Tappe, I.; Rapp,
U. R. Recent Results Cancer Res. 1997, 143, 237; Monia, B. P.;
Johnston, J. F.; Geiger, T.; Muller, M.; Fabbro, D. Nature Medicine
1996, 2, 668).
To support progressive tumor growth beyond the size of 1-2
mm.sup.3, it is recognized that tumor cells require a functional
stroma, a support structure consisting of fibroblast, smooth muscle
cells, endothelial cells, extracellular matrix proteins, and
soluble factors (Folkman, J., Semin Oncol, 2002. 29(6 Suppl 16),
15-8). Tumors induce the formation of stromal tissues through the
secretion of soluble growth factors such as PDGF and transforming
growth factor-beta (TGF-beta), which in turn stimulate the
secretion of complimentary factors by host cells such as fibroblast
growth factor (FGF), epidermal growth factor (EGF), and vascular
endothelial growth factor (VEGF). These stimulatory factors induce
the formation of new blood vessels, or angiogenesis, which brings
oxygen and nutrients to the tumor and allows it to grow and
provides a route for metastasis. It is believed some therapies
directed at inhibiting stroma formation will inhibit the growth of
epithelial tumors from a wide variety of Histological types.
(George, D. Semin Oncol, 2001. 28(5 Suppl 17), 27-33; Shaheen, R.
M., et al., Cancer Res, 2001. 61(4); 1464-8; Shaheen, R. M., et al.
Cancer Res, 1999. 59(21), 5412-6). However, because of the complex
nature and the multiple growth factors involved in angiogenesis
process and tumor progression, an agent targeting a single pathway
may have limited efficacy. It is desirable to provide treatment
against a number of key signaling pathways utilized by tumors to
induce angiogenesis in the host stroma. These include PDGF, a
potent stimulator of stroma formation (Ostman, A. and C. H. Heldin,
Adv Cancer Res, 2001, 80, 1-38), FGF, a chemo-attractant and
mitogen for fibroblasts and endothelial cells, and VEGF, a potent
regulator of vascularization.
PDGF is another key regulator of stromal formation which is
secreted by many tumors in a paracrine fashion and is believed to
promote the growth of fibroblasts, smooth muscle and endothelial
cells, promoting stroma formation and angiogenesis. PDGF was
originally identified as the v-sis oncogene product of the simian
sarcoma virus (Heldin, C. H., et al., J Cell Sci Suppl, 1985, 3,
65-76). The growth factor is made up of two peptide chains,
referred to as A or B chains which share 60% homology in their
primary amino acid sequence. The chains are disulfide cross linked
to form the 30 kDa mature protein composed of either AA, BB or AB
homo- or heterodimers. PDGF is found at high levels in platelets,
and is expressed by endothelial cells and vascular smooth muscle
cells. In addition, the production of PDGF is up regulated under
low oxygen conditions such as those found in poorly vascularized
tumor tissue (Kourembanas, S., et al., Kidney Int, 1997, 51(2),
438-43). PDGF binds with high affinity to the PDGF receptor, a 1106
amino acid 124 kDa transmembrane tyrosine kinase receptor (Heldin,
C. H., A. Ostman, and L. Ronnstrand, Biochim Biophys Acta, 1998.
1378(1), 79-113). PDGFR is found as homo- or heterodimer chains
which have 30% homology overall in their amino acid sequence and
64% homology between their kinase domains (Heldin, C. H., et al.
Embo J, 1988, 7(5), 1387-93). PDGFR is a member of a family of
tyrosine kinase receptors with split kinase domains that includes
VEGFR2 (KDR), VEGFR3 (Flt4), c-Kit, and FLT3. The PDGF receptor is
expressed primarily on fibroblast, smooth muscle cells, and
pericytes and to a lesser extent on neurons, kidney mesangial,
Leydig, and Schwann cells of the central nervous system. Upon
binding to the receptor, PDGF induces receptor dimerization and
undergoes auto- and trans-phosphorylation of tyrosine residues
which increase the receptors' kinase activity and promotes the
recruitment of downstream effectors through the activation of SH2
protein binding domains. A number of signaling molecules form
complexes with activated PDGFR including PI-3-kinase, phospholipase
C-gamma, src and GAP (GTPase activating protein for p21-ras)
(Soskic, V., et al. Biochemistry, 1999, 38(6), 1757-64). Through
the activation of PI-3-kinase, PDGF activates the Rho signaling
pathway inducing cell motility and migration, and through the
activation of GAP, induces mitogenesis through the activation of
p21-ras and the MAPK signaling pathway.
In adults, it is believed the major function of PDGF is to
facilitate and increase the rate of wound healing and to maintain
blood vessel homeostasis (Baker, E. A. and D. J. Leaper, Wound
Repair Regen, 2000. 8(5), 392-8; Vu, J., A. Moon, and H. R. Kim,
Biochem Biophys Res Commun, 2001. 282(3), 697-700). PDGF is found
at high concentrations in platelets and is a potent chemoattractant
for fibroblast, smooth muscle cells, neutrophils and macrophages.
In addition to its role in wound healing PDGF is known to help
maintain vascular homeostasis. During the development of new blood
vessels, PDGF recruits pericytes and smooth muscle cells that are
needed for the structural integrity of the vessels. PDGF is thought
to play a similar role during tumor neovascularization. As part of
its role in angiogenesis PDGF controls interstitial fluid pressure,
regulating the permeability of vessels through its regulation of
the interaction between connective tissue cells and the
extracellular matrix. Inhibiting PDGFR activity can lower
interstitial pressure and facilitate the influx of cytotoxics into
tumors improving the anti-tumor efficacy of these agents (Pietras,
K., et al. Cancer Res, 2002. 62(19), 5476-84; Pietras, K., et al.
Cancer Res, to 2001. 61(7), 2929-34).
PDGF can promote tumor growth through either the paracrine or
autocrine stimulation of PDGFR receptors on stromal cells or tumor
cells directly, or through the amplification of the receptor or
activation of the receptor by recombination. Over expressed PDGF
can transform human melanoma cells and keratinocytes (Forsberg, K.,
et al. Proc Natl Aced Sci USA., 1993. 90(2), 393-7; Skobe, M. and
N. E. Fusenig, Proc Natl Aced Sci USA, 1998. 95(3), 1050-5), two
cell types that do not express PDGF receptors, presumably by the
direct effect of PDGF on stroma formation and induction of
angiogenesis. This paracrine stimulation of tumor stroma is also
observed in carcinomas of the colon, lung, breast, and prostate
(Bhardwaj, B., et al. Clin Cancer Res, 1996, 2(4), 773-82;
Nakanishi, K., et al. Mod Pathol, 1997, 10(4), 341-7; Sundberg, C.,
et al. Am J Pathol, 1997, 151(2), 479-92; Lindmark, G., et al. Lab
Invest, 1993, 69(6), 682-9; Vignaud, J. M., et al, Cancer Res,
1994, 54(20), 5455-63) where the tumors express PDGF, but not the
receptor. The autocrine stimulation of tumor cell growth, where a
large faction of tumors analyzed express both the ligand PDGF and
the receptor, has been reported in glioblastomas (Fleming, T. P.,
et al. Cancer Res, 1992, 52(16), 4550-3), soft tissue sarcomas
(Wang, J., M. D. Coltrera, and A. M. Gown, Cancer Res, 1994, 54(2),
560-4) and cancers of the ovary (Henriksen, R., et al. Cancer Res,
1993, 53(19), 4550-4), prostate (Fudge, K., C. Y. Wang, and M. E.
Stearns, Mod Pathol, 1994, 7(5), 549-54), pancreas (Funa, K., et
al. Cancer Res, 1990, 50(3), 748-53) and lung (Antoniades, H. N.,
et al., Proc Natl Acad Sci USA, 1992, 89(9), 3942-6). Ligand
independent activation of the receptor is found to a lesser extent
but has been reported in chronic myelomonocytic leukemia (CMML)
where the a chromosomal translocation event forms a fusion protein
between the Ets-like transcription factor TEL and the PDGF
receptor. In addition, activating mutations in PDGFR have been
found in gastrointestinal stromal tumors in which c-Kit activation
is not involved (Heinrich, M. C., et al., Science, 2003, 9, 9).
Certain PDGFR inhibitors will interfere with tumor stromal
development and are believed to inhibit tumor growth and
metastasis.
Another major regulator of angiogenesis and vasculogenesis in both
embryonic development and some angiogenic-dependent diseases is
vascular endothelial growth factor (VEGF; also called vascular
permeability factor, VPF). VEGF represents a family of isoforms of
mitogens existing in homodimeric forms due to alternative RNA
splicing. The VEGF isoforms are reported to be highly specific for
vascular endothelial cells (for reviews, see: Farrara et al.
Endocr. Rev. 1992, 93, 18; Neufield et al. FASEB J. 1999, 13,
9).
VEGF expression is reported to be induced by hypoxia (Shweiki et
al. Nature 1992, 359, 843), as well as by a variety of cytokines
and growth factors, such as interleukin-1, interleukin-6, epidermal
growth factor and transforming growth factor. To date, VEGF and the
VEGF family members have been reported to bind to one or more of
three transmembrane receptor tyrosine kinases (Mustonen et al. J.
Cell Biol., 1995, 129, 895), VEGF receptor-1 (also known as flt-1
(fms-like tyrosine kinase-1)), VEGFR-2 (also known as kinase insert
domain containing receptor (KDR); the murine analogue of KDR is
known as fetal liver kinase-1 (flk-1)), and VEGFR-3 (also known as
flt-4). KDR and flt-1 have been shown to have different signal
transduction properties (Waltenberger et al. J. Biol. Chem. 1994,
269, 26988); Park et al. Oncogene 1995, 10, 135). Thus, KDR
undergoes strong ligand-dependant tyrosine phosphorylation in
intact cells, whereas flt-1 displays a weak response. Thus, binding
to KDR is believed to be a critical requirement for induction of
the full spectrum of VEGF-mediated biological responses.
In vivo, VEGF plays a central role in vasculogenesis, and induces
angiogenesis and permeabilization of blood vessels. Deregulated
VEGF expression contributes to the development of a number of
diseases that are characterized by abnormal angiogenesis and/or
hyperpermeability processes. It is believed regulation of the
VEGF-mediated signal transduction cascade by some agents can
provide a useful mode for control of abnormal angiogenesis and/or
hyperpermeability processes.
Angiogenesis is regarded as an important prerequisite for growth of
tumors beyond about 1-2 mm. Oxygen and nutrients may be supplied to
cells in tumors smaller than this limit through diffusion. However,
it is believed every tumor is dependent on angiogenesis for
continued growth after it has reached a certain size. Tumorigenic
cells within hypoxic regions of tumors respond by stimulation of
VEGF production, which triggers activation of quiescent endothelial
cells to stimulate new blood vessel formation. (Shweiki et al.
Proc. Nat'l. Acad. Sci., 1995, 92, 768). In addition, VEGF
production in tumor regions where there is no angiogenesis may
proceed through the ras signal transduction pathway (Grugel et al.
J. Biol. Chem., 1995, 270, 25915; Rak et al. Cancer Res. 1995, 55,
4575). In situ hybridization studies have demonstrated VEGF mRNA is
strongly upregulated in a wide variety of human tumors, including
lung (Mattern et al. Br. J. Cancer 1996, 73, 931), thyroid
(Viglietto et al. Oncogene 1995, 11, 1569), breast (Brown et al.
Human Pathol. 1995, 26, 86), gastrointestinal tract (Brown et al.
Cancer Res. 1993, 53, 4727; Suzuki et al. Cancer Res. 1996, 56,
3004), kidney and bladder (Brown et al. Am. J. Pathol. 1993, 143I,
1255), ovary (Olson et al. Cancer Res. 1994, 54, 1255), and
cervical (Guidi et al. J. Nat'l Cancer Inst. 1995, 87, 12137)
carcinomas, as well as angiosarcoma (Hashimoto et al. Lab. Invest.
1995; 73, 859) and several intracranial tumors (Plate et al. Nature
1992, 359, 845; Phillips et al. Int. J. Oncol. 1993, 2, 913;
Berkman et al. J. Clin. Invest., 1993, 91, 153). Neutralizing
monoclonal antibodies to KDR have been shown to be efficacious in
blocking tumor angiogenesis (Kim et al. Nature 1993, 362, 841;
Rockwell et al. Mol. Cell. Differ. 1995, 3, 315).
Over expression of VEGF, for example under conditions of extreme
hypoxia, can lead to intraocular angiogenesis, resulting in
hyperproliferation of blood vessels, leading eventually to
blindness. Such a cascade of events has been observed for a number
of retinopathies, including diabetic retinopathy, ischemic
retinal-vein occlusion, and retinopathy of prematurity (Aiello et
al. New Engl. J. Med. 1994, 331, 1480; Peer et al. Lab. Invest.
1995, 72, 638), and age-related macular degeneration (AMD; see,
Lopez et al. Invest. Opththalmol. Vis. Sci. 1996, 37, 855).
In rheumatoid arthritis (RA), the in-growth of vascular pannus may
be mediated by production of angiogenic factors. Levels of
immunoreactive VEGF are high in the synovial fluid of RA patients,
while VEGF levels were low in the synovial fluid of patients with
other forms of arthritis of with degenerative joint disease (Koch
et al. J. Immunol. 1994, 152, 4149). The angiogenesis inhibitor
AGM-170 has been shown to prevent neovascularization of the joint
in the rat collagen arthritis model (Peacock et al. J. Exper. Med.
1992, 175, 1135).
Increased VEGF expression has also been shown in psoriatic skin, as
well as bullous disorders associated with subepidermal blister
formation, such as bullous pemphigoid, erythema multiforme, and
dermatitis herpetiformis (Brown et al. J. Invest. Dermatol. 1995,
104, 744).
The vascular endothelial growth factors (VEGF, VEGF-C, VEGF-D) and
their receptors (VEGFR2, VEGFR3) are not only key regulators of
tumor angiogenesis, but also lymphangiogenesis. VEGF, VEGF-C and
VEGF-D are expressed in most tumors, primarily during periods of
tumor growth and, often at substantially increased levels. VEGF
expression is stimulated by hypoxia, cytokines, oncogenes such as
ras, or by inactivation of tumor suppressor genes (McMahon, G.
Oncologist 2000, 5(Suppl. 1), 3-10; McDonald, N. Q.; Hendrickson,
W. A. Cell 1993, 73, 421-424)
The biological activities of the VEGFs are mediated through binding
to their receptors. VEGFR3 (also called Flt-4) is predominantly
expressed on lymphatic endothelium in normal adult tissues. VEGFR3
function is needed for new lymphatic vessel formation, but not for
maintenance of the pre-existing lymphatics. VEGFR3 is also
upregulated on blood vessel endothelium in tumors. Recently VEGF-C
and VEGF-D, ligands for VEGFR3, have been identified as regulators
of lymphangiogenesis in mammals. Lymphangiogenesis induced by
tumor-associated lymphangiogenic factors could promote the growth
of new vessels into the tumor, providing tumor cells access to
systemic circulation. Cells that invade the lymphatics could find
their way into the bloodstream via the thoracic duct. Tumor
expression studies have allowed a direct comparison of VEGF-C,
VEGF-D and VEGFR3 expression with clinicopathological factors that
relate directly to the ability of primary tumors to spread (e.g.,
lymph node involvement, lymphatic invasion, secondary metastases,
and disease-free survival). In many instances, these studies
demonstrate a statistical correlation between the expression of
lymphangiogenic factors and the ability of a primary solid tumor to
metastasize (Skobe, M. et al, Nature Med. 2001, 7(2), 192-198;
Stacker, S. A. et al. Nature Med. 2001, 7(2), 186-191; Makinen, T.
et al. Nature Med. 2001, 7(2), 199-205; Mandriota, S. J. et al.
EMBO J. 2001, 20(4), 672-82; Karpanen, T. et al. Cancer Res. 2001,
61(5), 1786-90; Kubo, H. et al. Blood 2000, 96(2), 546-53).
Hypoxia appears to be an important stimulus for VEGF production in
malignant cells. Activation of p38 MAP kinase is required for VEGF
induction by tumor cells in response to hypoxia (Blaschke, F. et
al. Biochem. Biophys. Res. Commun. 2002, 296, 890-896; Shemirani,
B. et al. Oral Oncology 2002, 38, 251-257). In addition to its
involvement in angiogenesis through regulation of VEGF secretion,
p38 MAP kinase promotes malignant cell invasion, and migration of
different tumor types through regulation of collagenase activity
and urokinase plasminogen activator expression (Laferriere, J. et
al. J. Biol. Chem. 2001, 276, 33762-33772; Westermarck, J. et al.
Cancer Res. 2000, 60, 7156-7162; Huang, S. et al. J. Biol. Chem.
2000, 275, 12266-12272; Simon, C. et al. Exp. Cell Res. 2001, 271,
344-355).
Some diarylureas have been described as having activity as
serine-threonine kinase and/or as tyrosine kinase inhibitors. The
utility of these diarylureas as an active ingredient in
pharmaceutical compositions for the treatment of cancer,
angiogenesis disorders, and inflammatory disorders has been
demonstrated. See Redman et al., Bioorg. Med. Chem. Lett. 2001, 11,
9-12; Smith et al., Bioorg. Med. Chem. Lett. 2001, 11, 2775-2778;
Dumas et al., Bioorg. Med. Chem. Lett. 2000, 10, 2047-2050; Dumas
et al., Bioorg. Med. Chem. Lett. 2000, 10, 2051-2054; Ranges et
al., Book of Abstracts, 220.sup.th ACS National Meeting,
Washington, D.C., USA, MEDI 149; Dumas et al., Bioorg. Med. Chem.
Lett. 2002, 12, 1559-1562; Lowinger et al., Clin. Cancer Res. 2000,
6(suppl.), 335; Lyons et al., Endocr.-Relat. Cancer 2001, 8,
219-225; Riedl et al., Book of Abstracts, 92.sup.nd AACR Meeting;
New Orleans, La., USA, abstract 4956; Khire et al., Book of
Abstracts, 93.sup.rd AACR Meeting, San Francisco, Calif., USA,
abstract 4211; Lowinger et al., Curr. Pharm. Design 2002, 8,
99-110; Regan et al., J. to Med. Chem. 2002, 45, 2994-3008;
Pargellis et al., Nature Struct. Biol. 2002, 9(4), 268-272; Carter
et al., Book of Abstracts, 92.sup.nd AACR Meeting, New Orleans,
La., USA, abstract 4954; Vincent et al., Book Of Abstracts,
38.sup.th ASCO Meeting, Orlando, Fla., USA, abstract 1900; Hilger
et al., Book of Abstracts, 38.sup.th ASCO Meeting, Orlando, Fla.,
USA, abstract 1916; Moore et al., Book of Abstracts, 38.sup.th ASCO
Meeting, Orlando, Fla., USA, abstract 1816; Strumberg et al., Book
of Abstracts, 38.sup.th ASCO Meeting, Orlando, Fla., USA, abstract
121; Madwed J B: Book of Abstracts, Protein Kinases Novel Target
Identification and Validation for Therapeutic Development, San
Diego, Calif., USA, March 2002; Roberts et al., Book of Abstracts,
38.sup.th ASCO Meeting, Orlando, Fla., USA, abstract 473; Tolcher
et al., Book of Abstracts, 38.sup.th ASCO Meeting, Orlando, Fla.,
USA, abstract 334; and Karp et al., Book of Abstracts, 38.sup.th
AACR Meeting, San Francisco, Calif., USA, abstract 2753.
Despite the advancements in the art, there remains a need for
cancer treatments and anti-cancer compounds.
DESCRIPTION OF THE INVENTION
The present invention pertains to: (i) novel compounds of formula
(I) below, salts, metabolites, prodrugs and diastereoisomeric forms
thereof (both isolated stereoisomers and mixtures of
stereoisomers), collectively referred to herein as the "compounds
of the invention"; (ii) pharmaceutical compositions containing
compounds of this invention; and (iii) use of compounds of this
invention or pharmaceutical compositions containing compounds of
this invention for treating diseases, e.g., hyper-proliferative and
angiogenesis disorders, as a sole agent or in combination with
other anti-cancer agents.
Formula I is as follows:
##STR00002## A is pyrimidine, optionally substituted with 1 to 3
substituents which are independently R.sup.1, OR.sup.1,
S(O).sub.pR.sup.1, C(O)R.sup.1, C(O)OR.sup.1, C(O)NR.sup.1R.sup.2,
halogen, hydroxy, amino, cyano, or nitro; B is phenyl, naphthyl, or
pyridyl, optionally substituted with 1 to 4 substituents which are
independently C.sub.1-C.sub.5 linear or branched alkyl,
C.sub.1-C.sub.5 linear or branched haloalkyl, C.sub.1-C.sub.3
alkoxy, hydroxy, amino, C.sub.1-C.sub.3 alkylamino, C.sub.1-C.sub.6
dialkylamino, halogen, cyano, or nitro. B is preferably phenyl,
optionally substituted with 1-4 substituents which are
independently C.sub.1-C.sub.5 linear or branched alkyl,
C.sub.1-C.sub.5 linear or branched haloalkyl, C.sub.1-C.sub.3
alkoxy, hydroxy, amino, C.sub.1-C.sub.3 alkylamino, C.sub.1-C.sub.6
dialkylamino, halogen, cyano, or nitro.
L is a bridging group which is: (a)
--(CH.sub.2).sub.m--O--(CH.sub.2).sub.l--, (b)
--(CH.sub.2).sub.m--(CH.sub.2).sub.l--, (c)
--(CH.sub.2).sub.m--C(O)--(CH.sub.2).sub.l, (d)
--(CH.sub.2).sub.m--NR.sup.3--(CH.sub.2).sub.l--, (e)
--(CH.sub.2).sub.m--NR.sup.3C(O)--(CH.sub.2).sub.l--, (f)
--(CH.sub.2).sub.m--S--(CH.sub.2).sub.l--, or (g)
--(CH.sub.2).sub.m--C(O)NR.sup.3--(CH.sub.2).sub.l--.
The integers m and l are independently selected from 0-4 and are
typically selected from 0-2. The group
--(CH.sub.2).sub.m--(CH.sub.2).sub.l-- defines a single bond where
m and l are 0.
L is most preferably --O-- or --S--.
M is a pyridine or pyrimidine ring, optionally substituted with 1-3
substituents which are independently selected from: (1)
C.sub.1-C.sub.5 linear or branched alkyl; (2) C.sub.1-C.sub.5
linear or branched haloalkyl; (3) C.sub.1-C.sub.3 alkoxy; (4)
hydroxy; (5) amino; (6) C.sub.1-C.sub.3 alkylamino; (7)
C.sub.1-C.sub.6 dialkylamino; (8) halogen; (9) nitro; (10)
C(O)NR.sup.4R.sup.5; (11) C(O)OR.sup.4; (12) C(O)R.sup.4; (13) CN;
(14) C(S)NR.sup.4R.sup.5; (15a) C(O)NR.sup.7--NR.sup.4R.sup.5;
(15b) C(O)NR.sup.7--R.sup.4C(O)NR.sup.4R.sup.5; (16) tetrazolyl;
(17) imidazolyl; (18) imidazoline-2-yl; (19)
1,3,4-oxadiazoline-2-yl; (20) 1,3-thiazoline-2-yl; (21)
5-thioxo-4,5-dihydro-1,3,4-thiazoline-2-yl; (22)
5-oxo-4,5-dihydro-1,3,4-oxadiazoline-2-yl; or (23) a group of the
formula
##STR00003##
M is preferably pyridine, optionally substituted with 1-3
substituents which are independently selected from the groups (1)
to (13) cited above.
Each of R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 is
independently (a) hydrogen, (b) C.sub.1-C.sub.5 linear, branched,
or cyclic alkyl, (c) phenyl, (d) C.sub.1-C.sub.3 phenyl-alkyl, (e)
up to per-halo substituted C.sub.1-C.sub.5 linear or branched
alkyl, or (f) --(CH.sub.2).sub.q--X.
The substituent X is a 5 or 6 membered heterocyclic ring,
containing at least one atom selected from oxygen, nitrogen and
sulfur, which is saturated, partially saturated, or aromatic, or a
8-10 membered bicyclic heteroaryl having 1-4 heteroatoms selected
from the group consisting of O, N and S.
In addition, R.sup.4 and R.sup.5 taken together may form a 5 or 6
membered aliphatic ring, which may be interrupted by an atom
selected from N, O or S. This is optionally substituted with 1-3
substituents which are independently C.sub.1-C.sub.5 linear or
branched alkyl, up to perhalo substituted C.sub.1-C.sub.5 linear or
branched alkyl, C.sub.1-C.sub.3 alkoxy, hydroxy, oxo, carboxy,
amino, C.sub.1-C.sub.3 alkylamino, C.sub.1-C.sub.6 dialkylamino,
halogen, cyano, or nitro.
R.sup.6 is independently:
(a) hydrogen, (b) C.sub.1-C.sub.5 linear, branched, or cyclic
alkyl, (c) cyano, (d) nitro, (e) up to per-halo substituted
C.sub.1-C.sub.5 linear or branched alkyl. or (f) --C(O)R.sup.7,
where R.sup.7 is C.sub.1-C.sub.5 linear, branched, or cyclic alkyl.
R.sup.6 is preferably independently: (a) hydrogen, (b)
C.sub.1-C.sub.5 linear, branched, or cyclic alkyl, or (c) cyano or
(d) nitro, and most preferably, R.sup.6 is independently: (a)
hydrogen, (b) C.sub.1-C.sub.5 linear, branched, or cyclic alkyl, or
(c) cyano.
R.sup.7 is hydrogen, or C.sub.1-C.sub.5 linear, branched, or cyclic
alkyl.
The variable q is an integer 0, 1, 2, 3, or 4. The variable p is an
integer 0, 1, or 2. When any moiety is "substituted", it can have
up to the highest number of indicated substituents, and each
substituent can be located at any available position on the moiety
and can be attached through any available atom on the substituent.
"Any available position" means any position on the moiety that is
chemically accessible through means known in the art or taught
herein and that does not create an unduly unstable molecule. When
there are two or more substituents on any moiety, each substituent
is defined independently of any other substituent and can,
accordingly, be the same or different.
The term "optionally substituted" means that the moiety so modified
may be either unsubstituted, or substituted with the identified
substituent(s).
It is understood that since M is pyridine, the term "hydroxy" as a
pyridine substituent includes 2-, 3-, and 4-hydroxypyridine, but
also includes those structures referred to in the art as
1-oxo-pyridine, 1-hydroxy-pyridine and pyridine N-oxide.
Where the plural form of the word compounds, salts, and the like,
is used herein, this is taken to mean also a single compound, salt,
or the like.
The term "C.sub.1-C.sub.5alkyl", as used herein, means straight or
branched chain alkyl groups having from one to five carbon atoms,
which may be linear or branched with single or multiple branching.
Such groups include methyl, ethyl, n-propyl, isopropyl, n-butyl,
isobutyl, sec-butyl, tert-butyl, and the like.
The term "C.sub.1-C.sub.5 haloalkyl", as used herein, means a
saturated hydrocarbon radical having up to five carbon atoms, which
is substituted with a least one halogen atom, up to perhalo. The
radical may be linear or branched with single or multiple
branching. The halo substituent(s) include fluoro, chloro, bromo,
or iodo. Fluoro, chloro and bromo are preferred, and fluoro and
chloro are more preferred. The halogen substituent(s) can be
located on any available carbon. When more than one halogen
substituent is present on this moiety, they may be the same or
different. Examples of such halogenated alkyl substituents include
but are not limited to chloromethyl, dichloromethyl,
trichloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl,
2,2,2-trifluoroethyl, and 1,1,2,2-tetrafluoroethyl, and the
like.
The term "C.sub.1-C.sub.3 alkoxy", as used herein, means a straight
or branched chain alkoxy group having from one to three saturated
carbon atoms which may be linear or branched with single or
multiple branching, and includes such groups as methoxy, ethoxy,
n-propoxy, isopropoxy, and the like. It also includes halogenated
groups such as 2,2-dichloroethoxy, trifluoromethoxy, and the
like.
Halo or halogen means fluoro, chloro, bromo, or iodo. Fluoro,
chloro and bromo are preferred, and fluoro and chloro are more
preferred.
The term "C.sub.1-C.sub.3alkylamine", as used herein, means
methylamino, ethylamino, propylamino or isopropylamino.
Examples of C.sub.1-C.sub.6 dialkylamine include but are not
limited to diethylamino, ethyl-isopropylamino, methyl-isobutylamino
and dihexylamino.
The term "heteroaryl", as used herein, refers to both monocyclic
and bicyclic heteroaryl rings. Monocyclic heteroaryl means an
aromatic monocyclic rings having 5 to 6 ring atoms, at least one of
which is a hetero atom selected from N, O and S, the remaining
atoms being carbon. When more than one hetero atom is present in
the moiety, they are selected independently from the other(s) so
that they May be the same or different. Monocyclic heteroaryl rings
include, but are not limited to pyrrole, furan, thiophene,
imidazole, pyrazole, thiazole, oxazole, isoxazole, isothiazole,
triazole, tetrazole, thiadiazole, oxadiazole, pyridine, pyrimidine,
pyridazine, pyrazine, and triazine.
The term "bicyclic heteroaryl", as used herein, means fused
bicyclic moieties where one of the rings is chosen from the
monocyclic heteroaryl rings described above and the second ring is
either benzene or another monocyclic heteroaryl ring described
above. When both rings in the bicyclic moiety are heteroaryl rings,
they may be the same or different, as long as they are chemically
accessible by means known in the art. Bicyclic heteroaryl rings
include synthetically accessible 5-5, 5-6, or 6-6 fused bicyclic
aromatic structures including, for example but not by way of
limitation, benzoxazole (fused benzene and oxazole), indazole
(fused benzene and pyrazole), quinoline (fused phenyl and
pyridine), quinazoline (fused pyrimidine and benzene),
imidazopyrimidine (fused imidazole and pyrimidine), naphthyridine
(two fused pyridines), and the like.
The term "5 or 6 membered heterocyclic ring, containing at least
one atom selected from oxygen, nitrogen and sulfur, which is
saturated, partially saturated, or aromatic" includes, by no way of
limitation, tetrahydropyrane, tetrahydrofurane, 1,3-dioxolane,
1,4-dioxane, morpholine; thiomorpholine, piperazine, piperidine,
piperidinone, tetrahydropyrimidone, pentamethylene sulfide,
tetramethylene sulfide, dihydropyrane, dihydrofurane,
dihydrothiophene, pyrrole, furan, thiophene, imidazole, pyrazole,
thiazole, oxazole, isoxazole, Isothiazole, triazole, pyridine,
pyrimidine, pyridazine, pyrazine, triazine, and the like.
Non-limiting examples of group of the formula
##STR00004## where R.sup.4 and R.sup.5 taken together may form a 5
or 6 membered aliphatic ring, which may be interrupted by an atom
selected from N, O or S, which is optionally substituted
include:
##STR00005##
The term "C.sub.1-C.sub.3 phenyl-alkyl" includes, by no way of
limitation, 3-phenyl-propyl, phenyl-1-methyl-ethyl. Substituted
examples include 2-[2-chlorophenyl]ethyl,
3,4-dimethylphenyl-methyl, and the like.
The compounds of Formula I may contain one or more asymmetric
centers, depending upon the location and nature of the various
substituents desired.
Asymmetric carbon atoms may be present in the (R) or (S)
configuration or (R,S) configuration. In certain instances,
asymmetry may also be present due to restricted rotation about a
given bond, for example, the central bond adjoining two substituted
aromatic rings of the specified compounds. Substituents on a ring
may also be present in either cis or trans form. It is intended
that all such configurations (including enantiomers and
diastereomers), are included within the scope of the present
invention. Preferred compounds are those with the absolute
configuration of the compound of Formula I which produces the more
desirable biological activity. Separated, pure or partially
purified isomers or racemic mixtures of the compounds of this
invention are also included within the scope of the present
invention. The purification of said isomers and the separation of
said isomeric mixtures can be accomplished by standard techniques
known in the art.
The optical isomers can be obtained by resolution of the racemic
mixtures according to conventional processes, for example, by the
formation of diastereoisomeric salts is using an optically active
acid or base or formation of covalent diastereomers. Examples of
appropriate acids are tartaric, diacetyltartaric, ditoluoyltartaric
and camphorsulfonic acid. Mixtures of diastereoisomers can be
separated into their individual diastereomers on the basis of their
physical and/or chemical differences by methods known in the art,
for example, by chromatography or fractional crystallization. The
optically active bases or acids are then liberated from the
separated diastereomeric salts. A different process for separation
of optical isomers involves the use of chiral chromatography (e.g.,
chiral HPLC columns), with or without conventional derivation,
optimally chosen to maximize the separation of the enantiomers.
Suitable chiral HPLC columns are manufactured by Diacel, e.g.,
Chiracel OD and Chiracel OJ among many others, all routinely
selectable. Enzymatic separations, with or without derivitization,
are also useful. The optically active compounds of Formula I can
likewise be obtained by chiral syntheses utilizing optically active
starting materials.
The present invention also relates to useful forms of the compounds
as disclosed herein, such as pharmaceutically acceptable salts,
metabolites and prodrugs of all the compounds Formula (I). The term
"pharmaceutically acceptable salt" refers to a relatively
non-toxic, inorganic or organic acid addition salt of a compound of
the present invention. For example, see S. M: Berge, et al.
"Pharmaceutical Salts," J. Pharm. Sci. 1977, 66, 1-19.
Pharmaceutically acceptable salts include those obtained by
reacting the main compound, functioning as a base, with an
inorganic or organic acid to form a salt, for example, salts of
hydrochloric acid, sulfuric acid, phosphoric acid, methane sulfonic
acid, camphor sulfonic acid, oxalic acid, maleic acid, succinic
acid and citric acid. Pharmaceutically acceptable salts also
include those in which the main compound functions as an acid and
is reacted with an to appropriate base to form, e.g., sodium,
potassium, calcium, magnesium, ammonium, and choline salts. Those
skilled in the art will further recognize that acid addition salts
of the claimed compounds may be prepared by reaction of the
compounds with the appropriate inorganic or organic acid via any of
a number of known methods. Alternatively, alkali and alkaline earth
metal salts are prepared by reacting the compounds of the invention
with the appropriate base via a variety of known methods.
Representative salts of the compounds of this invention include the
conventional non-toxic salts and the quaternary ammonium salts
which are formed, for example, from inorganic or organic acids or
bases by means well known in the art. For example, such acid
addition salts include acetate, adipate, alginate, ascorbate,
aspartate, benzoate, benzenesulfonate, bisulfate, butyrate,
citrate, camphorate, camphorsulfonate, cinnamate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate,
hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,
hydroiodide, 2-hydroxyethanesulfonate, itaconate, lactate, maleate,
mandelate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oxalate, palmoate, pectinate, persulfate,
3-phenylpropionate, picrate, pivalate, propionate, succinate,
sulfonate, tartrate, thiocyanate, tosylate, and undecanoate.
Base salts include alkali metal salts such as potassium and sodium
salts, alkaline earth metal salts such as calcium and magnesium
salts, and ammonium salts with organic bases such as
dicyclohexylamine and N-methyl-D-glucamine. Additionally, basic
nitrogen containing groups may be quaternized with such agents as
lower alkyl halides such as methyl, ethyl, propyl, and butyl
chlorides, bromides and iodides; dialkyl sulfates like dimethyl,
diethyl, and dibutyl sulfate; and diamyl sulfates, long chain
halides such as decyl, lauryl, myristyl and stearyl chlorides,
bromides and iodides, aralkyl halides like benzyl and phenethyl
bromides and others.
Certain compounds of this invention can be further modified with
labile functional groups that are cleaved after in vivo
administration to furnish the parent active agent to and the
pharmacologically inactive derivatizing (functional) group. These
derivatives, commonly referred to as prodrugs, can be used, for
example, to alter the physicochemical properties of the active
agent, to target the active agent to a specific tissue, to alter
the pharmacokinetic and pharmacodynamic properties of the active
agent, and to reduce undesirable side effects
Prodrugs of the invention include, e.g., the esters of appropriate
compounds of this invention are well-tolerated, pharmaceutically
acceptable esters such as alkyl esters including methyl, ethyl,
propyl, isopropyl, butyl, isobutyl or pentyl esters. Additional
esters such as phenyl-C.sub.1-C.sub.5 alkyl may be used, although
methyl ester is preferred.
Methods for synthesizing prodrugs are described in the following
reviews on the subject, which are incorporated herein by reference
for their description of these methods: Higuchi, T.; Stella, V.
eds. Prodrugs As Novel Drug Delivery Systems. ACS Symposium Series.
American Chemical Society: Washington, D.C. (1975). Roche, E. B.
Design of Biopharmaceutical Properties through Prodrugs and
Analogs. American Pharmaceutical Association: Washington, D.C.
(1977). Sinkula, A. A.; Yalkowsky, S. H. J Pharm Sci. 1975, 64,
181-210. Stella, V. J.; Charman, W. N. Naringrekar, V. H. Drugs
1985, 29, 455-473. Bundgaard, H., ed. Design of Prodrugs. Elsevier:
New York (1985). Stella, V. J.; Himmelstein, K. J. J. Med. Chem.
1980, 23, 1275-1282. Han, H-K; Amidon, G. L. AAPS Pharmsci 2000, 2,
1-11. Denny, W. A. Eur. J. Med. Chem. 2001, 36, 577-595. Wermuth,
C. G. in Wermuth, C. G. ed. The Practice of Medicinal Chemistry
Academic Press: San Diego (1996), 697-715. Balant, L. P.; Doelker,
E. in Wolff, M. E. ed. Burgers Medicinal Chemistry And Drug
Discovery John Wiley & Sons: New York (1997), 949-982.
The metabolites of the compounds of this invention include oxidized
derivatives of the compounds of Formula I, wherein one or more of
the nitrogens are substituted with a hydroxy group; which includes
derivatives where the nitrogen atom of the pyridine group is in the
oxide form, referred to in the art as 1-oxo-pyridine or has a
hydroxy substituent, referred to in the art as
1-hydroxy-pyridine.
Compounds of interest within the scope of formula I are of formula
(III) including the salts, metabolites, prodrugs and
diastereoisomeric forms thereof,
##STR00006## wherein Ra is R.sup.1, OR.sup.1 or cyano; and B, L and
M are as defined above.
Another group of compounds of interest within the scope of formula
I are of formula (IV) including the salts, metabolites, prodrugs
and diastereoisomeric forms thereof,
##STR00007## wherein Ra is R.sup.1, OR.sup.1 or cyano; each Rc is
independently hydrogen, halogen, C.sub.1-C.sub.3 linear or branched
alkyl, C.sub.1-C.sub.3 linear or branched haloalkyl,
C.sub.1-C.sub.3 alkoxy or hydroxy; and L and M are as defined
above.
Another group of compounds of interest within the scope of formula
I are of formula (V) including the salts, metabolites, prodrugs and
diastereoisomeric forms thereof,
##STR00008## wherein Ra, Rc and M is are as defined above. In
groups of interest M is pyridine in formulae III, IV and V, and is
typically substituted by C(O)NR.sup.4R.sup.5 or CN. In certain
groups of interest, C(O)NR.sup.4R.sup.5 is C(O)NHCH.sub.3 or
C(O)NH.sub.2.
A further group of compounds of interest are of formula (II)
including the salts, metabolites; prodrugs and diastereoisomeric
forms thereof,
##STR00009## wherein Ra; Rc are as defined above and Rb is (1)
C.sub.1-C.sub.5 linear or branched alkyl; (2) C.sub.1-C.sub.5
linear or branched haloalkyl; (3) C.sub.1-C.sub.3 alkoxy; (4)
hydroxy; (5) amino; (6) C.sub.1-C.sub.3 alkylamino; (7)
C.sub.1-C.sub.6 dialkylamino; (8) halogen; (9) nitro; (10)
C(O)NR.sup.4R.sup.5; (11) C(O)OR.sup.4; (12) C(O)R.sup.4; (13) CN;
(14) C(S)NR.sup.4R.sup.5; (15)
C(O)NR.sup.7--R.sup.4C(O)NR.sup.4R.sup.5; or (16) hydrogen; with
each of R.sup.1, R.sup.4, R.sup.5 and R.sup.7 independently as
defined above.
For a group of compounds of Formula II of interest, Rb is
C.sub.1-C.sub.5 linear or branched alkyl; C.sub.1-C.sub.3 alkoxy;
halogen; C(O)NR.sup.4R.sup.5; CN; C(S)NR.sup.4R.sup.5 or
C(O)NR.sup.7--R.sup.4C(O)NR.sup.4R.sup.5. For another group of
compounds of formula II of interest, Rb is C.sub.1-C.sub.5 linear
or branched alkyl; halogen; C(O)NR.sup.4R.sup.5 or CN. In a further
sub-groups, Rb is C(O)NR.sup.4R.sup.5 or CN or Rb is only
C(O)NR.sup.4R.sup.5.
For the compounds of formula II and the groups thereof mentioned
above, there are sub-groups where each Rc, independently, is
hydrogen or fluorine and Rb is C(O)NHCH.sub.3 or C(O)NH.sub.2.
General Preparative Methods
The particular process to be utilized in the preparation of the
compounds used in this embodiment of the invention depends upon the
specific compound desired. Such factors as the selection of the
specific substituents play a role in the path to be followed in the
preparation of the specific compounds of this invention. Those
factors are readily recognized by one of ordinary skill in the
art.
The compounds of the invention may be prepared by use of known
chemical reactions and procedures. Nevertheless, the following
general preparative methods are presented to aid the reader in
synthesizing the compounds of the present invention, with more
detailed particular examples being presented below in the
experimental section describing the working examples.
All variable groups of these methods are as described in the
generic description if they are not specifically defined below.
When a variable group or substituent with a given symbol is used
more than once in a given structure, it is to be understood that
each of these groups or substituents may be independently varied
within the range of definitions for that symbol. It is recognized
that compounds of the invention with each claimed optional
functional group cannot be prepared with each of the below-listed
methods. Within the scope of each method optional substituents are
used which are stable to the reaction conditions, or the functional
groups which may participate in the reactions are present in
protected form where necessary, and the removal of such protective
groups is completed at appropriate stages by methods well known to
those skilled in the art.
The compounds of the invention can be made according to
conventional chemical methods, and/or as disclosed below, from
starting materials which are either commercially available or
producible according to routine, conventional chemical methods.
General methods for the preparation of the compounds are given
below, and the preparation of representative compounds is
specifically illustrated in examples.
##STR00010##
The compounds (I) can be synthesized according to the reaction
sequence shown in the General Method 1 above. Thus, the compounds
(I) can be synthesized by reacting amino compounds (III) with
isocyanate compounds (II).
The compounds (II) are commercially available or can be synthesized
according to methods commonly known to those skilled in the art,
e.g. from treatment of an amine with phosgene or a phosgene
equivalent such as trichloromethyl chloroformate (diphosgene),
bis(trichloromethyl)carbonate (triphosgene), or
N,N'-carbonyldiimidazole (CDI); or, alternatively by a Curtius-type
rearrangement of an amide, or a carboxylic acid derivative, such as
an ester, an acid halide or an anhydride. The compounds (III) are
commercially available or can be synthesized according methods
commonly known to those skilled in the art.
Alternatively, compounds of Formula (I) can be prepared according
to general method 2, where aminopyrimidines of formula (IV) and
amino compounds of formula (III) are coupled together to form a
urea of Formula (I), with the use of a coupling agent such as
carbonyldiimidazole, phosgene, diphosgene, triphosgene, and the
like. The coupling step may be performed in an inert solvent such
as dioxane, diethylether, dichloromethane, chloroform,
tetrahydrofuran, toluene, and the like, at a temperature selected
between 0.degree. C. and reflux. This coupling may be achieved
using these reagents alone, or, in the presence of an organic or
inorganic base as described in the art.
##STR00011##
In addition specific preparations of diaryl ureas are already
described in the patent literature, and can be adapted to the
compounds of the present invention. For example, Miller S. et al,
"Inhibition of p38 Kinase using Symmetrical and Unsymmetrical
Diphenyl Ureas" PCT Int. Appl. WO 99 32463, Miller, S et al.
"Inhibition of raf Kinase using Symmetrical and Unsymmetrical
Substituted Diphenyl Ureas" PCT Int. Appl., WO 99 32436, Dumas, J.
et al., "Inhibition of p38 Kinase Activity using Substituted
Heterocyclic Ureas" PCT Int. Appl., WO 99 32111, Dumas, J. et al.,
"Method for the Treatment of Neoplasm by Inhibition of raf Kinase
using N-Heteroaryl-N'-(hetero)arylureas" PCT Int. Appl., WO 99
32106, Dumas, J. et al., "Inhibition of p38 Kinase Activity using
Aryl- and Heteroaryl-Substituted Heterocyclic Ureas" PCT Int.
Appl., WO 99 32110, Dumas, J., et al., "Inhibition of raf Kinase
using Aryl- and Heteroaryl-Substituted Heterocyclic Ureas" PCT Int.
Appl., WO 99 32455, Riedl, B., et al., "O-Carboxy Aryl Substituted
Diphenyl Ureas as raf Kinase Inhibitors" PCT Int. Appl., WO 00
42012, Riedl, B., et al., "O-Carboxy Aryl Substituted Diphenyl
Ureas as p38 Kinase Inhibitors" PCT Int. Appl., WO 00 41698, Dumas,
J. et al. "Heteroaryl ureas containing nitrogen hetero-atoms as p38
kinase inhibitors" U.S. Pat. Appl. Publ., US 20020065296, Dumas, J.
et al. "Preparation of N-aryl-N'-[(acylphenoxy)phenyl]ureas as raf
kinase inhibitors" PCT Int. Appl., WO 02 62763, Dumas, J. et al.
"inhibition of raf kinase using quinolyl, isoquinolyl or pyridyl
ureas" PCT Int. Appl., WO 02 85857, Dumas, J. et al. "Preparation
of quinolyl, isoquinolyl or pyridyl-ureas as inhibitors of raf
kinase for the treatment of tumors and/or cancerous cell growth"
U.S. Pat. Appl. Publ., US 20020165394. All the preceding patent
applications are hereby incorporated by reference.
The reaction of the compounds (II) with (III) is carried out
preferably in a solvent. Suitable solvents comprise the customary
organic solvents which are inert under the reaction conditions.
Non-limiting examples include ethers such as diethyl ether,
dioxane, tetrahydrofuran, 1,2-dimethoxy ethane; hydrocarbons such
as benzene, toluene, xylene, hexane, cyclohexane, mineral oil
fractions; halogenated hydrocarbons such as dichloromethane,
trichloromethane, carbon tetrachloride, dichloroethane,
trichloroethylene, chlorobenzene; alcohols such as methanol,
ethanol, n-propanol, isopropanol; esters such as ethyl acetate;
ketones such as acetone; nitrites such as acetonitrile;
heteroaromatics such as pyridine; polar solvents such as dimethyl
formamide and hexamethyl phosphoric acid tris-amide; and mixtures
of the above-mentioned solvents. Toluene, benzene, and
dichloromethane are preferred.
The compounds (III) are generally employed in an amount of from 1
to 3 mol per mol of compounds (II); an equimolar amount or slight
excess of compounds (III) is preferred.
The reaction of the compounds (II) with (III) is generally carried
out within a relatively wide temperature range. In general, they
are carried out in a range of from -20 to 200.degree. C.,
preferably from 0 to 100.degree. C., and more preferably from 25 to
50.degree. C. The steps of this reaction are generally carried out
under atmospheric pressure. However, it is also possible to carry
them out under superatmospheric pressure or under reduced pressure
(for example, in a range of from 0.5 to 5 bar). The reaction time
can generally be varied within a relatively wide range. In general,
the reaction is finished after a period of from 2 to 24 hours,
preferably from 6 to 12 hours.
Synthetic transformations that may be employed in the synthesis of
compounds of Formula I and in the synthesis of intermediates
involved in the synthesis of compounds of Formula I are known by or
accessible to one skilled in the art. Collections of synthetic
transformations may be found in compilations, such as: J. March.
Advanced Organic Chemistry, 4th ed.; John Wiley: New York (1992) R.
C. Larock. Comprehensive Organic Transformations, 2nd ed.;
Wiley-VCH: New York (1999) F. A. Carey; R. J. Sundberg. Advanced
Organic Chemistry, 2nd ed.; Plenum Press: New York (1984) T. W.
Greene; P. G. M. Wuts. Protective Groups in Organic Synthesis, 3rd
ed.; John Wiley: New York (1999) L. S. Hegedus. Transition Metals
in the Synthesis of Complex Organic Molecules, 2nd ed., University
Science Books: Mill Valley, Calif. (1994) L. A. Paquette, Ed. The
Encyclopedia of Reagents for Organic Synthesis; John Wiley: New
York (1994) A. R. Katritzky; O. Meth-Cohn; C. W. Rees, Eds.
Comprehensive Organic Functional Group Transformations; Pergamon
Press: Oxford, UK (1995) G. Wilkinson; F. G A. Stone; E. W. Abel,
Eds. Comprehensive Organometallic Chemistry; Pergamon Press:
Oxford, UK (1982) B. M. Trost; I. Fleming. Comprehensive Organic
Synthesis; Pergamon Press: Oxford, UK (1991) A. R. Katritzky; C. W.
Rees Eds. Comprehensive Heterocylic Chemistry; Pergamon Press:
Oxford, UK (1984) A. R. Katritzky; C. W. Rees; E. F. V. Scriven,
Eds. Comprehensive Heterocylic Chemistry II; Pergamon Press:
Oxford, UK (1996) C. Hansch; P. G. Sammes; J. B. Taylor, Eds.
Comprehensive Medicinal Chemistry: Pergamon Press: Oxford, UK
(1990).
In addition, recurring reviews of synthetic methodology and related
topics include Organic Reactions; John Wiley: New York; Organic
Syntheses; John Wiley: New York; Reagents for Organic Synthesis:
John Wiley: New York; The Total Synthesis of Natural Products; John
Wiley: New York; The Organic Chemistry of Drug Synthesis; John
Wiley New York; Annual Reports in Organic Synthesis; Academic
Press: San Diego Calif.; and Methoden der Organischen Chemie
(Houben-Weyl); Thieme: Stuttgart, Germany. Furthermore, databases
of synthetic transformations include Chemical Abstracts, which may
be searched using either CAS OnLine or SciFinder, Handbuch der
Organischen Chemie (Beilstein), which may be searched using
SpotFire, and REACCS.
The preparation of the compounds of the present invention is
further illustrated in Examples 1-13.
Compositions of the Compounds of this Invention
This invention also relates to pharmaceutical compositions
containing one or more compounds of the present invention. These
compositions can be utilized to achieve the desired pharmacological
effect by administration to a patient in need thereof. A patient,
for the purpose of this invention, is a mammal, including a human,
in need of treatment for the particular condition or disease.
The active compounds of the present invention can act systemically
and/or locally. For this purpose, they can be administered in a
suitable manner, such as for example by oral, parenteral,
pulmonary, nasal, sublingual, lingual, buccal, rectal, dermal,
transdermal, conjunctival or aural administration or in the form of
an implant or stent. The active compound can be administered in
forms suitable for these modes of administration.
Suitable forms of oral administration are those according to the
prior art which function by releasing the active compound rapidly
and/or in a modified or controlled manner and which contain the
active compound in a crystalline and/or amorphous and/or dissolved
form, such as for example tablets (which are uncoated or coated,
for example with enteric coatings or coatings which dissolve after
a delay in time or insoluble coatings which control the release of
the active compound), tablets or films/wafers which disintegrate
rapidly in the oral cavity or films/lyophilisates, capsules (e.g.
hard or soft gelatin capsules), dragees, pellets, powders,
emulsions, suspensions and solutions.
Parenteral administration can be carried out by avoiding an
absorption step (e.g. by intravenous, intraarterial, intracardial,
intraspinal or intralumbar administration) or by including
absorption (e.g. by intramuscular, subcutaneous, intracutaneous or
intraperitoneal administration). Suitable parenteral administration
forms are for example injection and infusion formulations in the
form of solutions, suspensions, emulsions, lyophilisates and
sterile powders.
Suitable forms of administration for the other modes of
administration are for example inhalation devices (such as for
example powder inhalers, nebulizers), nasal drops, solutions and
sprays; tablets or films/wafers for lingual, sublingual or buccal
administration or capsules, suppositories, ear and eye
preparations, vaginal capsules, aqueous suspensions (lotions or
shaking mixtures), lipophilic suspensions, ointments, creams,
transdermal or therapeutic systems, milky lotions, pastes, foams,
dusting powders, implants or stents.
The active compounds can be converted into the above mentioned
forms of administration in a manner known to the skilled man and in
accordance with the prior art using inert, non-toxic,
pharmaceutically suitable auxiliaries. The latter include for
example excipients (e.g. microcrystalline cellulose, lactose,
mannitol, etc.), solvents (e.g. liquid polyethylene glycols),
emulsifiers and dispersants or wetting agents (e.g. sodium dodecyl
sulphate, polyoxysorbitan oleate etc.), binders (e.g. polyvinyl
pyrrolidone), synthetic and/or natural polymers (e.g. albumin),
stabilizers (e.g. antioxidants, such as, for example, ascorbic
acid), dyes (e.g. inorganic pigments such as iron oxides) or taste-
and/or odor-corrective agents.
Method of Treating Hyper-proliferative Disorders
The present invention also relates to a method for using the
compounds of Formula I and pharmaceutical compositions containing
them to treat mammalian hyper-proliferative disorders, including
cancer. The term "hyper-proliferative disorders" and/or "cancer"
not only refers to solid tumors, such as cancers of the breast,
respiratory tract, brain, reproductive organs, digestive tract,
urinary tract, eye, liver, skin, head and neck, thyroid,
parathyroid and their distant metastases, but also includes
lymphomas, sarcomas, and leukemias.
Examples of breast cancer include, but are not limited to invasive
ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in
situ, and lobular carcinoma in situ.
Examples of cancers of the respiratory tract include, but are not
limited to small-cell and non-small-cell lung carcinoma, as well as
bronchial adenoma and pleuropulmonary blastoma.
Examples of brain cancers include, but are not limited to brain
stem and hypophthalmic glioma, cerebellar and cerebral astrocytoma,
medulloblastoma, ependymoma, as well as neuroectodermal and pineal
tumor.
Tumors of the male reproductive organs include, but are not limited
to prostate and testicular cancer. Tumors of the female
reproductive organs include, but are not limited to endometrial,
cervical, ovarian, vaginal, and vulvar cancer, as well as sarcoma
of the uterus.
Tumors of the digestive tract include, but are not limited to anal,
colon, colorectal, esophageal, gallbladder, gastric, pancreatic,
rectal, small intestine, and salivary gland cancers.
Tumors of the urinary tract include, but are not limited to
bladder, penile, kidney, renal pelvis, ureter, and urethral
cancers.
Eye cancers include, but are not limited to intraocular melanoma
and retinoblastoma.
Examples of liver cancers include, but are not limited to
hepatocellular carcinoma (liver cell carcinomas with or without
fibrolamellar variant), cholangiocarcinoma (intrahepatic bile duct
carcinoma), and mixed hepatocellular cholangiocarcinoma.
Skin cancers include, but are not limited to squamous cell
carcinoma, Kaposi's sarcoma, malignant melanoma, Merkel cell skin
cancer, and non-melanoma skin cancer.
Head-and-neck cancers include, but are not limited to
laryngeal/hypopharyngeal/nasopharyngeal/oropharyngeal cancer, and
lip and oral cavity cancer.
Lymphomas include, but are not limited to AIDS-related lymphoma,
non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, Hodgkin's
disease, and lymphoma of the central nervous system.
Sarcomas include, but are not limited to sarcoma of the soft
tissue, fibrosarcoma, osteosarcoma, malignant fibrous histiocytoma,
lymphosarcoma, and rhabdomyosarcoma.
Leukemias include, but are not limited to acute myeloid leukemia,
acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic
myelogenous leukemia, and hairy cell leukemia.
These disorders have been well characterized in humans, but also
exist with a similar etiology in other mammals, and can be treated
by administering the compounds and pharmaceutical compositions of
the present invention.
Any raf or VEGFR polypeptide can be modulated in accordance with
present invention, including both wild-type and mutant forms. Raf
or raf-1 kinase is a family of serine/threonine kinases which
comprise at least three family members, A-Raf, B-Raf, and c-raf or
Raf-1. See, e.g., Dhillon and Kolch, Arch. Biochem. Biophys.,
404:3-9, 2002. C-raf and B-Raf are preferred targets for compounds
of the present invention. Activating B-Raf mutations (e.g., V599E
mutant) have been identified in various cancers, including
melanoma, and the compounds described herein can be utilized to
inhibit their activity. Mutations, include mutations in K-RAS;
mutations in the BRAF gene, such as mutations at position 599, such
as V599E, and/or positions 461, 462, 463, 465, 468, 593, 596, 60,
etc., which are associated with cancers, such as melanoma.
VEGFR-2, as indicated above, plays a role in angiogenesis, and
therefore inhibiting it is useful to treat tumors and other
diseases associated with neovasculature, including rheumatoid
arthritis, osteoarthritis, asthma, pulmonary fibrosis, age-related
macular degeneration (ARMD), diabetic retinopathy, macular
degeneration, and retinopathy of prematurity (ROP), endometriosis,
cancer, Coats' disease, peripheral retinal neovascularization,
neovascular glaucoma, psoriasis, retrolental fibroplasias,
angiofibroma, inflammation, etc.
Methods of the present invention include modulating tumor cell
proliferation, including inhibiting cell proliferation. The latter
indicates that the growth and/or differentiation of tumor cells is
reduced, decreased, diminished, slowed, etc. The term
"proliferation" includes any process which relates to cell growth
and division, and includes differentiation and apoptosis. As
discussed above, raf kinases play a key role in the activation of
the cytoplasmic signaling cascade involved in cell proliferation,
differentiation, and apoptosis. For example, studies have found
that inhibiting c-raf-1 by anti-sense oligonucleotides can block
cell proliferation (see above). Any amount of inhibition is
considered therapeutic.
Additionally, the present invention relates to methods of screening
patients to determine their susceptibility to compounds of the
present invention. For example, the presenting invention relates to
methods of selecting subjects having a disease for treatment with a
compound of formula I, comprising, one or more of the following
steps in any effective order, e.g., measuring the expression or
activity of Raf, VEGFR-2, or other kinase receptors, in a sample
obtained from a subject having a disease, and administering said
compound of formula I to subjects who are identified as having high
levels of expression or activity, where said compound is a compound
of formula I of claim 1.
The term "susceptibility" is used broadly to indicate, e.g.,
ability to respond, toxicity or other adverse effects, etc. For
example, the invention relates to methods of determining whether a
condition can be modulated by a compound disclosed herein,
comprising measuring the expression or activity of Raf, VEGFR-2, or
other kinase proteins in cells having said condition. The results
can be used to determine or predict whether a subject will respond
to a compound of the present invention. For example, where the
condition is a tumor, the methods can be used to predict whether
the tumor is susceptible to compounds of the present invention. By
the term "susceptible," it is meant that tumor can be treated with
it, e.g., causing tumor regression or cell death, inhibiting cell
proliferation, inhibiting tumor growth, inhibiting tumor
metastasis, etc.
Whether a condition, such as a tumor, is susceptible to a compound
of the present invention can be determined routinely. For instance,
cells or tissues (e.g., tumor cells, a biopsy sample, etc.) that
exhibit the condition can be assayed for the presence and/or
activity of Raf, VEGFR-2, or other kinase proteins. When high
levels of expression and/or activity are identified, this can
indicate that the subject will respond to, and benefit from, a
compound of the present invention. Levels of gene expression (e.g.,
mRNA levels), gene amplification, or gene product activity (e.g.,
tyrosine kinase activity) can be utilized to characterize the state
of the cell with respect to the corresponding gene and signaling
pathway. For example, the target genes of the present invention
possess tyrosine kinase activity, and therefore kinase activity can
be used to assess the cell or tissue state. In the example below,
activity was measured by looking at the levels of substrate
phosphorylated by it. This can be done quantitatively (e.g., using
isotopes, spectroscopy, etc.) or semi-quantitatively as in the
example where the levels were assessed visually and assigned a
level of intensity from +1 to +4. A cell or tissue which has a high
level of phosphorylated substrate (and a high number of cells
exhibiting the heightened activity) can be considered to have a
high level of kinase activity, and therefore be a candidate for
therapy with a compound of the present invention. More than one
activity can be assessed, and the results from several targets can
be utilized in deciding whether a subject's condition (e.g., a
tumor) will be responsive to a compound of the present
invention.
High levels of target activity can be relative to a control or
other standard. For instance, in the example below, high levels of
activity were with reference to a cell type (stromal) in the tissue
section which normally does not express substantial levels of the
target gene. High levels can therefore be where cells express a
statistically higher amount of measured activity or phosphorylated
substrate than the standard or control used as a comparison. High
levels can also be where 25% or more cells express the target
activity.
The method can further comprise a step of comparing the expression
in a sample with a normal control, or expression in a sample
obtained from normal or unaffected tissue. Comparing can be done
manually, against a standard, in an electronic form (e.g., against
a database), etc. The normal control can be a standard sample that
is provided with the assay; it can be obtained from adjacent, but
unaffected, tissue from the same patient; or, it can be
pre-determined values, etc. Gene expression, protein expression
(e.g., abundance in a cell), protein activity (e.g., kinase
activity), etc., can be determined.
For instance, a biopsy from a cancer patient can be assayed for the
presence, quantity, and/or activity of Raf, VEGFR-2, or other
kinase proteins. Increased expression or activity of one or more of
these can indicate that the cancer can be targeted for treatment by
a compound of the present invention. For example, raf activity can
be monitored by its ability to initiate the cascade leading to ERK
phosphorylation (i.e., raf/MEK/ERK), resulting in phospho-ERK.
Increased phospho-ERK levels in a cancer shows that its raf
activity is elevated, suggesting the use of compounds of the
present invention to treat it. In addition to biopsy samples,
phospho-ERK (other markers) can also be measured in other body
fluids, such as serum, blood, cerebral spinal fluid, urine, etc.,
such as in peripheral blood lymphocytes (PBLs). For the latter,
inhibition of ERK phosphorylation can be measured following
activation with phorbol myristate acetate using antibodies as
described in the examples below.
In addition, patients having cancer can be selected and monitored
on the basis of whether the tissue is experiencing
neovascularization, and how much. This can be assessed as discussed
above, e.g., using immunohistochemistry for vessel markers (e.g.,
CD31), circulating levels of a VGFR ligand, etc.
Patient selection and monitoring can also be made on the basis of
the appearance in a body fluid (such as blood) above normal levels
of the shedded ectodomains derived from the various receptors,
including the extracellular portions of VEGFR-2 or other kinase
receptors. Detection methods can be carried out routinely, e.g.,
using antibodies which specifically bind to the extracellular
domain.
Measuring expression includes determining or detecting the amount
of the polypeptide present in a cell or shed by it, as well as
measuring the underlying mRNA, where the quantity of mRNA present
is considered to reflect the quantity of polypeptide manufactured
by the cell. Furthermore, the genes for Raf, VEGFR-2, and other
kinase proteins can be analyzed to determine whether there is a
gene defect responsible for aberrant expression or polypeptide
activity. Genes sequences are publically available; e.g.,
NM.sub.--004333 Homo sapiens v-raf murine sarcoma viral oncogene
homolog B1 (BRAF); NM.sub.--002253 Homo sapiens VEGFR2.
The present invention also provides methods of assessing the
efficacy of a compound of the present invention in treating a
disease, comprising one or more of the following steps in any
effective order, e.g., measuring the expression or activity of Raf,
VEGFR-2, or other kinase proteins in a sample obtained from said
subject who has been treated with a compound of the present
invention, and determining the effects of said compound on said
expression or activity. The measuring step can be carried out as
described already.
For instance, biopsy samples can be removed from patients who have
been treated with a compound of the present invention, and then
assayed for the presence and/or activity of the mentioned signaling
molecules. As discussed above, decreased levels of phospho-ERK in
the cancer tissue (e.g., compared to a normal tissue or before
treatment) indicate that the compound is exerting in vivo efficacy
and a therapeutic effect.
Determining the effects of the compound on expression or activity
includes performing a comparison step between a tissue sample and a
control, or other type of standard. Examples of standards that can
be used, include, but are not limited to, a tissue sample prior to
treatment, a tissue sample from an unaffected tissue or from an
unaffected region of the affected tissue (e.g., from a region of
the tissue which is not transformed, cancerous, etc.), etc. A
standard can also be a value, or range of values, that is
representative of normal levels of, expression that have been
established for that marker. The comparison can also be made
between samples collected from at least two different timepoints
during the treatment regimen with a compound of the present
invention. For example, samples can be collected from various times
after initiation of the drug treatment, and analysis of expression
and/or activity levels can be used to monitor the
progress/prognosis of the subject, e.g., how the subject is
responding to the drug regimen. Any timepoint can be used, e.g.,
daily, twice a week, weekly, every two weeks, every month, yearly,
a plurality of timepoints (at least 2, 3, 4, 8, 12, etc.).
The method can be used to determine appropriate dosages and dosing
is regimens, e.g., how much compound to administer and at what
frequency to administer it. By monitoring its effect on the
signaling molecules in the tissue, the clinician can determine the
appropriate treatment protocol and whether it is achieving the
desired effect, e.g., on modulating or inhibiting the signal
transduction pathway. For instance, if the compound is not
effective in knocking down the amounts of a marker, such as
phospho-ERK, the dosage can be increased in the patient or given
more frequently. Similarly, dosages and/or frequency can be reduced
when it is shown that the compound is effective in knocking down
the levels of phospho-ERK or other marker for the disease state.
Since the compounds can be administered in combination with others
treatments, e.g., radiation, chemotherapy, and other agents, the
monitoring of the subject can be used to assess the combined
effects of the treatment regimen on the progress of the
disease.
The total amount of the active ingredient (compounds of Formula I)
to be administered to a patient will generally range from about
0.01 mg/kg to about 50 mg/kg body weight per day. Based upon
standard laboratory techniques known to evaluate compounds useful
for the treatment of hyper-proliferative disorders, by standard
toxicity tests and by standard pharmacological assays for the
determination of treatment of the conditions identified above in
mammals, and by comparison of these results with the results of
known medicaments that are used to treat these conditions, the
effective dosage of the compounds and pharmaceutical compositions
of this invention can readily be determined by those skilled in the
art. The amount of the administered active ingredient can vary
widely according to such considerations as the particular compound
and dosage unit employed, the mode and time of administration, the
period of treatment, the age, sex, and general condition of the
patient treated, the nature and extent of the condition treated,
the rate of drug metabolism and excretion, the potential drug
combinations and drug-drug interactions, and the like.
The compounds and pharmaceutical compositions of this invention can
be administered as the sole agent or in combination with one or
more other therapies where the combination causes no unacceptable
adverse effects. For example, they can be combined with cytotoxic
agents, signal transduction inhibitors, or with other anti-cancer
agents or therapies, as well as with admixtures and combinations
thereof.
In one embodiment, the compounds and pharmaceutical compositions of
the present invention can be combined with cytotoxic anti-cancer
agents. Examples of such agents can be found in the 11.sup.th
Edition of the Merck Index (1996). These agents include, by no way
of limitation, asparaginase, bleomycin, carboplatin, carmustine,
chlorambucil, cisplatin, colaspase, cyclophosphamide, cytarabine,
dacarbazine, dactinomycin, daunorubicin, doxorubicin (adriamycine),
epirubicin, etoposide, 5-fluorouracil, hexamethylmelamine,
hydroxyurea, ifosfamide, irinotecan, leucovorin, lomustine,
mechlorethamine, 6-mercaptopurine, mesna, methotrexate, mitomycin
C, mitoxantrone, prednisolone, prednisone, procarbazine, raloxifen,
streptozocin, tamoxifen, thioguanine, topotecan, vinblastine,
vincristine, and vindesine.
Other cytotoxic drugs suitable for use with the compounds and
pharmaceutical compositions of the invention include, but are not
limited to, those compounds acknowledged to be used in the
treatment of neoplastic diseases in Goodman and Gilman's The
Pharmacological Basis of Therapeutics (Ninth Edition, 1996,
McGraw-Hill). These agents include, by no way of limitation,
aminoglutethimide, L-asparaginase, azathioprine, 5-azacytidine
cladribine, busulfan, diethylstilbestrol,
2',2'-difluorodeoxycytidine, docetaxel, erythrohydroxynonyladenine,
ethinyl estradiol, 5-fluorodeoxyuridine, 5-fluorodeoxyuridine
monophosphate, fludarabine phosphate, fluoxymesterone, flutamide,
hydroxyprogesterone caproate, idarubicin, interferon,
medroxyprogesterone acetate, megestrol acetate, melphalan,
mitotane, paclitaxel, pentostatin, N-phosphonoacetyl-L-aspartate
(PALA), plicamycin, semustine, teniposide, testosterone propionate,
thiotepa, trimethylmelamine, uridine, and vinorelbine.
Other cytotoxic anti-cancer agents suitable for use in combination
with the compounds and pharmaceutical compositions of the invention
also include newly discovered cytotoxic principles such as
oxaliplatin, gemcitabine, capecitabine, epothilone and its natural
or synthetic derivatives, temozolomide (Quinn et al., J. Clin.
Oncology 2003, 21(4), 646-651), tositumomab (Bexxar), trabedectin
(Vidal et al., Proceedings of the American Society for Clinical
Oncology 2004, 23, abstract 3181), and the inhibitors of the
kinesin spindle protein Eg5 (Wood et al., Curr. Opin. Pharmacol.
2001, 1, 370-377).
In another embodiment, the compounds and pharmaceutical
compositions of the present invention can be combined with other
signal transduction inhibitors. Of particular interest are signal
transduction inhibitors which target the EGFR family, such as EGFR,
HER-2, and HER-4 (Raymond et al., Drugs 2000, 60 (Suppl. 1), 15-23;
Harari et al., Oncogene 2000, 19 (53), 6102-6114), and their
respective ligands. Examples of such agents include, by no way of
limitation, antibody therapies such as Herceptin (trastuzumab),
Erbitux (cetuximab), and pertuzumab. Examples of such therapies
also include, by no way of limitation, small-molecule kinase
inhibitors such as ZD-1839/Iressa (Baselga et al., Drugs 2000, 60
(Suppl. 1), 33-40), OSI-774/Tarceva (Pollack et al. J. Pharm. Exp.
Ther. 1999, 291(2), 739-748), CI-1033 (Bridges, Curr. Med. Chem.
1999, 6, 825-843), GW-2016 (Lackey et al., 92.sup.nd AACR Meeting,
New Orleans, Mar. 24-28, 2001, abstract 4582), CP-724,714. Wall et
al., Proceedings of the American Society for Clinical Oncology
2004, 23, abstract 3122), HKI-272 (Rabindran et al., Cancer Res.
2004, 64, 3958-3965), and EKB-569 (Greenberger et al., 11.sup.th
NCI-EORTC-AACR Symposium on New Drugs in Cancer Therapy, Amsterdam,
Nov. 7-10, 2000, abstract 388).
In another embodiment, the compounds and pharmaceutical
compositions of the present invention can be combined with other
signal transduction inhibitors targeting receptor kinases of the
split-kinase domain families (VEGFR, FGFR, PDGFR, flt-3, c-kit,
c-fms, and the like), and their respective ligands. These agents
include, by no way of limitation, antibodies such as Avastin
(bevacizumab). These agents also include, by no way of limitation,
small-molecule inhibitors such as STI-571/Gleevec (Zvelebil, Curr.
Opin. Oncol., Endocr. Metab. Invest. Drugs 2000, 2(1), 74-82),
PTK-787 (Wood et al., Cancer Res. 2000, 60(8), 2178-2189), SU-11248
(Demetri et al., Proceedings of the American Society for Clinical
Oncology 2004, 23, abstract 3001), ZD-6474 (Hennequin et al.,
92.sup.nd AACR Meeting, New Orleans, Mar. 24-28, 2001, abstract
3152), AG-13736 (Herbst et al., Clin. Cancer Res. 2003, 9, 16
(suppl 1), abstract C253), KRN-951 (Taguchi et al., 95.sup.th
AACR-Meeting, Orlando, Fla., 2004, abstract 2575), CP-547,632
(Beebe et al., Cancer Res. 2003, 63, 7301-7309), CP-673,451
(Roberts et al., Proceedings of the American Association of Cancer
Research 2004, 45, abstract 3989), CHIR-258 (Lee et al.,
Proceedings of the American Association of Cancer Research 2004,
45, abstract 2130), MLN-518 (Shen et al., Blood 2003, 102, 11,
abstract 476), and AZD-2171 (Hennequin et al., Proceedings of the
American Association of Cancer Research 2004, 45, abstract
4539).
In another embodiment, the compounds and pharmaceutical
compositions of the present invention can be combined with
inhibitors of the Raf/MEK/ERK transduction pathway (Avruch et al.,
Recent Prog. Horm. Res. 2001, 56, 127-155), or the PKB (akt)
pathway (Lawlor et al., J. Cell Sci. 2001, 114, 2903-2910). These
include, by no way of limitation, PD-325901 (Sebolt-Leopold et al.,
Proceedings of the American Association of Cancer Research 2004,
45, abstract 4003), and ARRY-142886 (Wallace et al., Proceedings of
the American Association of Cancer Research 2004, 45, abstract
3891).
In another embodiment, the compounds and pharmaceutical
compositions of the present invention can be combined with
inhibitors of histone deacetylase. Examples of such agents include,
by no way of limitation, suberoylanilide hydroxamic acid (SAHAi),
LAQ-824 (Ottmann et al., Proceedings of the American Society for
Clinical Oncology 2004, 23, abstract 3024), LBH-589 (Beck et al.,
Proceedings of the American Society for Clinical Oncology 2004, 23,
abstract 3025), MS-275 (Ryan et al., Proceedings of the American
Association of Cancer Research 2004, 45, abstract 2452), and
FR-901228 (Piekarz et al., Proceedings of the American Society for
Clinical Oncology 2004, 23, abstract 3028).
In another embodiment, the compounds and pharmaceutical
compositions of the present invention can be combined with other
anti-cancer agents such as proteasome inhibitors, and m-TOR
inhibitors. These include, by no way of limitation, bortezomib
(Mackay et al., Proceedings of the American Society for Clinical
Oncology 2004, 23, Abstract 3109), and CCI-779 (Wu et al.,
Proceedings of the American Association of Cancer Research 2004,
45, abstract 3849).
Generally, the use of cytotoxic and/or cytostatic anti-cancer
agents in combination with the compounds or pharmaceutical
compositions of the present invention wilt serve to: (1) yield
better efficacy in reducing the growth of a tumor or even eliminate
the tumor as compared to administration of either agent alone, (2)
provide for the administration of lesser amounts of the
administered agents, (3) provide for a chemotherapeutic treatment
protocol that is well tolerated in the patient with fewer
deleterious pharmacological complications than observed with single
agent chemotherapies and certain other combined therapies, (4)
provide for treating a broader spectrum of different cancer types
in mammals, especially humans, (5) provide for a higher response
rate among treated patients, (6) provide for a longer survival time
among treated patients compared to standard chemotherapy
treatments, (7) provide a longer time for tumor progression, and/or
(8) yield efficacy and tolerability results at least as good as
those of the agents is used alone, compared to known instances
where other cancer agent combinations produce antagonistic
effects.
It is believed that one skilled in the art, using the preceding
information and information available in the art, can utilize the
present invention to its fullest extent.
It should be apparent to one of ordinary skill in the art that
changes and modifications can be made to this invention without
departing from the spirit or scope of the invention as it is set
forth herein.
All publications, applications and patents cited above and below
are incorporated herein by reference.
EXAMPLES
Abbreviations used in this specification DBU
1,8-diazabicyclo[5.4.0]undec-7-ene DMF N,N-dimethyl formamide DCM
Dichloromethane DCE 1,2-dichloroethane DMSO dimethyl sulphoxide
HPLC High pressure liquid chromatography MPLC Medium pressure
liquid chromatography LC-MS liquid chromatography-coupled mass
spectroscopy RT retention time MP melting point NMR nuclear
resonance spectroscopy TLC thin layer chromatography ES
electrospray DMA N,N-dimethylacetamide HRMS high resolution mass
spectroscopy CDI 1,1'-carbonyldiimidazole HOBT
1-hydroxybenzotriazole EDCI
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride TMSCl
Trimethylsilyl chloride m-CPBA 3-chloroperbenzoic acid HEPES
N-(2-hydroxyethyl)-piperazine-N'-(2-ethane sulphonic acid)
Tris/hydrochloric acid tris(hydroxymethyl)-aminomethane
hydrochloride .TM.Triton X-100.RTM.
tert.-octyl-phenoxypolyethoxyethanol, Rohm & Haas, USA
The yield percentages of the following examples refer to the
starting component which was used in the lowest molar amount.
LC-MS conditions: HPLC-electrospray mass spectra (HPLC ES-MS) were
obtained using a Gilson HPLC system equipped with two Gilson 306
pumps, a Gilson 215 Autosampler, a Gilson diode array detector, a
YMC Pro C-18 column (2.times.23 mm, 120 A), and a Micromass LCZ
single quadrupole mass spectrometer with z-spray electrospray
ionization. Spectra were scanned from 120-1000 amu over 2 seconds.
ELSD (Evaporative Light Scattering Detector) data was also acquired
as an analog channel. Gradient elution was used with Buffer A as 2%
acetonitrile in water with 0.02% TFA and Buffer B as 2% water in
Acetonitrile with 0.02% TFA at 1.5 mL/min. Samples were eluted as
follows: 90% A for 0.5 minutes ramped to 95% B over 3.5 minutes and
held at 95% B for 0.5 minutes and then the column is brought back
to initial conditions over 0.1 minutes. Total run time is 4.8
minutes.
Preparation of 6-Trifluoromethyl-pyrimidin-4-ylamine
##STR00012##
The procedure was derived from methods described in U.S. Pat. No.
5,756,275 and WO 02/38569. In a 250 mL round bottom flask,
6-trifluoromethyl-4-pyrimidinol (10 g, 60.9 mmol) was dissolved in
70 mL phosphorus oxychloride (0.73 mol). The solution was heated at
reflux for 7 h. The cooled reaction solution was then added
gradually to 200 mL 30% ammonium hydroxide, and the resulting
mixture was stirred overnight at room temperature. The reaction
mixture was extracted with ethyl acetate (2.times.100 mL), and the
combined extracts were dried (MgSO.sub.4) and evaporated in vacuo
to give 6-trifluoromethyl-pyrimidin-4-ylamine (1.4 g, yield 14%) as
a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.50 (s,
1 H), 7.60 (broad s, 2 H), 6.90 (s, 1 H); LC-MS m/z 164.1
[M+H].sup.+.
Example 1
4-{3-fluoro-4-[3-(6-trifluoromethylpyrimidin-4-yl)ureido]phenoxy}pyridine--
2-carboxylic acid methylamide
##STR00013##
In a 8-mL screw-cap vial, 4-amino-6-trifluoromethylpyrimidine (170
mg, 1.04 mmol) was added to a slurry of 1,1'-carbonyldiimidazole
(169 mg, 1.04 mmol) in dichloroethane (0.35 mL). The mixture was
heated at 60.degree. C. for 30 hours.
4-(4Amino-3-fluoro-phenoxy)-pyridine-2-carboxylic acid methylamide
(170 mg, 1.04 mmol) was then added and the mixture was heated
overnight at to 60.degree. C. The solvent was evaporated under
reduced pressure, and the solid residue was washed with methanol to
give the title product as a white solid (139 mg, yield 30%).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.6 (s, 1 H), 9.7 (s,
1 H), 9.0 (s, 1 H), 8.8 (s, 1 H), 8.5 (d, 1 H), 8.20 (m, 2 H), 7.40
(m, 2 H), 7.18 (d, 1 H), 7.10 (d, 1 H), 2.80 (s, 3 H). LC-MS m/z
451.3 [M+H].sup.+.
Example 2
4-{4-[3-(6-trifluoromethyl-pyrimidin-4-yl)-ureido]-phenoxy}-pyridine-2-car-
boxylic acid methylamide
##STR00014##
The title compound was prepared in the same manner as
4-{3-fluoro-4-[3-(6-trifluoromethylpyrimidin-4-yl)ureido]phenoxy}pyridine-
-2-carboxylic-acid methylamide, replacing
4-(4-amino-3-fluoro-phenoxy)pyridine-2-carboxylic acid methylamide
for 4-(4-aminophenoxy)pyridine-2-carboxylic acid methylamide. LC-MS
m/z 433.1 [M+H].sup.+; TLC Rf=0.8 (EtOAc).
Example 3
4-{4-[3-(6-trifluoromethylpyrimidin-4-yl)ureido]phenoxy}-2-methylpyridine
##STR00015##
The title compound was prepared in the same manner as
4-{3-fluoro-4-[3-(6-trifluoromethyl-pyrimidin-4-yl)-ureido]-phenoxy}-pyri-
dine-2-carboxylic-acid methyl-amide, replacing
4-(4-amino-3-fluorophenoxypyridine-2-carboxylic acid methylamide
for 4-(2-methylpyridin-4-yloxy)phenylamine. LC-MS m/z 389.9
[M+H].sup.+; TLC Rf=0.45 (EtOAc).
Example 4
1-[2-Fluoro-4-(2-methylpyridin-4-yloxy)phenyl]-3-(6-trifluoromethylpyrimid-
in-4-yl)urea
##STR00016##
To a solution of 2-fluoro-4-(2-methylpyridin-4-yloxy)phenylamine
(100 mg, 0.61 mmol) and N,N-diethylisopropylamine (0.13 mL, 0.74
mmol, 1.2 eq) in anhydrous THF (4 mL) was added triphenylphosphine
(67.3 mg, 0.23 mmol, 0.37 eq) in one portion. The reaction mixture
was stirred at 75.degree. C. After 2.5 h a solution of
4-amino-6-trifluoropyrimidine (133.8 mg, 0.61 mmol, 1.0 eq) in
anhydrous THF (2.5 mL) was added, and the reaction mixture was
stirred at 75.degree. C. for 16 h. The reaction mixture was then
partitioned between EtOAc and saturated aqueous sodium bicarbonate
solution. The organic layer was washed with water and brine, dried
over sodium sulfate, filtered, and evaporated under reduced
pressure. The crude oil was purified using medium pressure liquid
chromatography (Biotage), eluting with 75% EtOAc/hexane.
Crystallization from DCM/hexane afforded the title compound (60 mg,
24%) as a pale yellow solid. .sup.1H-NMR (DMSO-d.sub.6) .delta.
10.54 (broad s, 1H), 9.69 (broad s, 1H), 9.02 (s, 1H), 8.32 (d,
J=6.0 Hz, 1H), 8.19 to 8.12 (m, 2H), 7.28 (dd, J=11.7, 2.7 Hz, 1H),
7.02 (ddd, J=9.0, 3.0, 1.2 Hz, 1H), 6.80 (d, J=2.4 Hz, 1H), 6.475
(dd, J=6.0, 2.7 Hz, 1H), 2.40 (s, 3H); LC-MS m/z 408 [M+H].sup.+,
RT=2.30 min.
Example 5
1-(6-tert-Butylpyrimidin-4-yl)-3-[4-(2-cyanopyridin-4-yloxy)-2-fluoropheny-
l]urea
##STR00017##
The title compound was prepared in the same manner described for
1-[2-fluoro-4-(2-methylpyridin-4-yloxy)phenyl]-3-(6-trifluoromethylpyrimi-
din-4-yl)urea, replacing
2-fluoro-4-(2-methylpyridin-4-yloxy)phenylamine for
4-(pyridin-4-yloxy)-phenylamine. .sup.1H-NMR (DMSO-d.sub.6) .delta.
10.21 (broad s, 1H), 9.68 (broad s, 1H), 8.99 (s, 1H), 8.44 to 8.43
(broad s, 2H), 8.16 (d, J=1.2 Hz, 1H), 7.60 to 7.57 (m, 2H), 7.18
to 7.14 (m, 2H), 6.88 (dd, J=4.5, 1.5 Hz, 2H); LC-MS m/z 376
[M+H].sup.+, RT=2.17 min.
Example 6
1-(6-tert-Butylpyrimidin-4-yl)-3-[4-(2-cyanopyridin-4-yloxy)-2-fluoropheny-
l]urea
##STR00018##
The title compound was prepared in the same manner described for
1-[2-fluoro-4-(2-methylpyridin-4-yloxy)phenyl]-3-(6-trifluoromethylpyrimi-
din-4-yl)urea, replacing
2-fluoro-4-(2-methylpyridin-4-yloxy)phenylamine for
3-fluoro-4-(2-chloro-pyridin-4-yl-oxy)phenylamine. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 10.30 (broad s, 1H), 9.88 (broad s, 1H),
9.01 (s, 1H), 8.29 (d, J=6.0 Hz, 1H), 8.15 (d, J=1.2 Hz, 1H), 7.75
(dd, J=12.6, 2.7 Hz, 1H), 7.40 (t, J=8.7 Hz, 1H), 7.34 to 7.29 (m,
1H), 7.06 (d, J=2.4 Hz, 1H), 6.97 (dd, J=5.7, 2.4 Hz, 1H); LC-MS
m/z 428, [M+H].sup.+, RT=3.77 min.
Example 7
4-{3-fluoro-4-[3-(6-tert-butylpyrimidin-4-yl)ureido]phenoxy}pyridine-2-car-
boxylic acid methylamide
##STR00019##
A solution of 6-amino-4-t-butylpyrimidine (20.0 mg; 0.13 mmol),
triphosgene (14.52 mg; 0.05 mmol) and diisopropylethylamine (20.51
mg; 0.16 mmol) in THF (0.7 mL) was heated at 70.degree. C. for 4 h.
A solution of 4-(4-amino-3-fluoro-phenoxy)-pyridine-2-carboxylic
acid methylamide (34.5 mg; 0.13 mmol) in DMF (1.5 mL) was then
added and the reaction mixture was heated at 70.degree. C. for
another 8 h, then extracted between EtOAc and saturated aqueous
NaHCO.sub.3. The organic layer was dried and evaporated under
reduced pressure to give a crude oil that was purified via HPLC to
give the title compound (14 mg, 9%). .sup.1H-NMR (CD.sub.3OD)
.delta. 8.73 (s, 1H), 8.48 (d, J=4.0 Hz, 1H), 8.29 (t, J=4.0 Hz,
1H), 7.57 (d, J=4.0 Hz, 1H), 7.58-7.08 (m, 3H), 7.01 (s, 1H), 2.94
(s, 3H), 1.36 (s, 9H). LC-MS m/z 439 [M+H].sup.+.
Example 8
1-(6-tert-Butylpyrimidin-4-yl)-3-[4-(2-cyanopyridin-4-yloxy)-2-fluoropheny-
l]urea
##STR00020##
To a solution of 6-amino-4-tert-butylpyrimidine (150.0 mg; 0.99
mmol) in anhydrous 1,2-dichloroethane (1.9 mL) was added
1,1'-carbonyldi(1,2,4-triazole) (195.4 mg, 1.19 mmol, 1.2 eq), and
the reaction mixture was stirred at 65.degree. C. for 3 days. A
solution of 4-(4-amino-3-fluorophenoxy)pyridine-2-carbonitrile
(227.4 mg; 0.99 mmol, 1.0 eq) in anhydrous 1,2-dichloroethane (1.9
mL) was then added, and the reaction mixture was heated at
65.degree. C. for 5 h. The reaction was diluted with EtOAc, and the
organic layer was washed with water and brine, dried over sodium
sulfate, and evaporated under reduced pressure to give a crude oil.
Trituration from DCM afforded the title compound (211 mg, 52%) as a
white solid. .sup.1H-NMR (DMSO-d.sub.6) .delta. 10.37 (broad s,
1H), 10.08 (broad s, 1H), 8.75 (d, J=1.2 Hz, 1H), 8.59 (d, J=5.4
Hz, 1H), 8.25 (t, J=9.3 Hz, 1H), 7.73 (d, J=2.7 Hz, 1H), 7.59 (s,
1H), 7.37 (dd, J=12.0, 2.7 Hz, 1H), 7.24 (dd, J=5.4, 2.4 Hz, 1H),
7.08 (ddd, J=9.0, 2.7, 1.5 Hz, 1H), 1.26 (s, 9H); LC-MS m/z 407
[M+H].sup.+, RT=3.17 min.
Example 9
4-{4-[3-(6-tert-Butylpyrimidin-4-yl)ureido]-3-fluorophenoxy}pyridine-2-car-
boxylic acid amide
##STR00021##
A mixture of
1-(6-tert-butylpyrimidin-4-yl)-3-[4-(2-cyanopyridin-4-yloxy)-2-fluoro-phe-
nyl]urea (141 mg, 0.35 mmol) and sodium percarbonate (with 25%
H.sub.2O.sub.2) (218 mg, 1.4 mmol, 4.0 eq) in 2:1 v/v acetone/water
(11 mL) was stirred at 60.degree. C. for 16 h. The reaction was
partitioned between ethyl acetate and water, and the aqueous layer
was extracted with ethyl acetate (2.times.100 mL). The combined
organic layers were washed with water and brine, dried over sodium
sulfate, filtered, and evaporated under reduced pressure.
Trituration from methanol afforded the title compound (63 mg, 43%)
as a white solid. .sup.1H-NMR (DMSO-d.sub.6) 10.32 (broad s, 1H),
10.08 (broad s, 1H), 8.75 (d, J=1.2 Hz, 1H), 8.51 (d, J=5.4 Hz,
1H), 8.23 (t, J=9.3 Hz, 1H), 8.12 (broad s, 1H), 7.71 (broad s,
1H), 7.61 (d, J=1.2 Hz, 1H), 7.40 (d, J=2.4 Hz, 1H), 7.36 (dd,
J=11.4, 2.4 Hz, 1H), 7.18 (dd, J=5.7, 2.7 Hz, 1H), 7.08 (ddd,
J=9.0, 2.7, 1.5 Hz, 1H), 1.26 (s, 9H); LC-MS m/z 425 [M+H].sup.+,
RT=3.16 min.
Example 10
4-{4-[3-(6-tert-Butylpyrimidin-4-yl)ureido]phenoxy}pyridine-2-carbothioic
acid amide
##STR00022##
The title compound was prepared in the same manner as described for
1-(6-tert-butylpyrimidin-4-yl)-3-[4-(2-cyanopyridin-4-yloxy)-2-fluorophen-
yl]urea, replacing
4-(4-amino-3-fluorophenoxy)pyridine-2-car-bonitrile for
4-(4-aminophenoxy)pyridine-2-thioamide. .sup.1H-NMR (DMSO-d.sub.6)
.delta. 10.32 (broad s, 1H), 10.20 (broad s, 1H), 10.13 (s, 1H),
9.93 (broad s, 1H), 9.72 (s, 1H), 8.74 (d, J=1.2 Hz, 1H), 8.46 (d,
J=5.7 Hz, 1H), 7.95 (d, J=2.1 Hz, 1H), 7.67 (d, J=1.2 Hz, 1H), 7.64
to 7.60 (m, 2H), 7.21 to 7.17 (m, 2H), 7.12 (dd, J=5.7, 3.0 Hz,
1H), 1.26 (s, 9H); LC-MS m/z 423 [M+H].sup.+, RT=3.39 min.
Example 11
1-(6-tert-Butylpyrimidin-4-yl)-3-[4-(2-cyanopyridin-4-yloxy)-2-fluoropheny-
l]urea
##STR00023##
The title compound was prepared in the same manner as described for
1-(6-tert-butylpyrimidin-4-yl)-3-[4-(2-cyanopyridin-4-yloxy)-2-fluorophen-
yl]urea, replacing
4-(4-amino-3-fluorophenoxy)pyridine-2-carbonitrile for
4-(4-aminophenoxy)pyridine-2-carboxylic acid
methylcarbamoyl-methylamide. .sup.1H-NMR (DMSO-d.sub.6) 10.10
(broad s, 1H), 9.71 (broad s, 1H), 8.87 (t, J=6.0 Hz, 1H), 8.73 (d,
J=1.5 Hz, 1H), 8.52 (d, J=5.4 Hz, 1H), 7.85 to 7.80 (m, 1H), 7.67
(d, J=1.2 Hz, 1H), 7.63 to 7.60 (m, 2H), 7.35 (d, J=2.4 Hz, 1H),
7.19 to 7.15 (m, 3H), 3.82 (d, J=6.0 Hz, 2H), 2.55 (d, J=4.2 Hz,
3H), 1.25 (s, 9H); LC-MS m/z 478 [M+H].sup.+, RT=2.47 min.
Example 12
4-{3-fluoro-4-[3-(6-methoxypyrimidin-4-yl)ureido]phenoxy}pyridine-2-carbox-
ylic acid methylamide
##STR00024##
A solution of 6-amino-4-methoxypyrimidine (50.0 mg; 0.39 mmol),
triphosgene (43.0 mg; 0.14 mmol) and diisopropylethylamine (60.7
mg; 0.47 mmol) in THF (2.0 mL) was heated at 70.degree. C. for 4 h.
A solution of 4-(4-amino-3-fluoro-phenoxy)-pyridine-2-carboxylic
acid methylamide (102.3 mg; 0.39 mmol) in DMF (1.0 mL) was then
added, and the reaction mixture was heated at 70.degree. C. for
another 8 h, then extracted between EtOAc and saturated aqueous
NaHCO.sub.3. The organic layer was dried and evaporated to give a
crude oil that was purified via HPLC to give the title compound (14
mg, 9%). .sup.1H-NMR (CD.sub.3OD) .delta. 8.36 (s, 1H), 8.35 (s,
1H), 8.14 (t, J=8.8 Hz, 1H), 7.46 (d, J=2.4 Hz, 1H), 7.00-6.86 (m,
3H), 6.60 (s, 1H), 3.86 (s, 3H), 2.83 (s, 3H); LC-MS m/z 413
[M+H].sup.+.
Example 13
4-{4-[3-(6-Phenylpyrimidin-4-yl)ureido]phenoxy}pyridine-2-carbothioic
acid amide
##STR00025##
The title compound was prepared in the same manner as described for
4-{-4-[3-(6-tert-butylpyrimidin-4-yl)ureido]phenoxy}pyridine-2-carbothioi-
c acid amide, replacing 6-amino-4-tert-butylpyrimidine for
6-amino-4-phenylpyrimidine. .sup.1H-NMR (DMSO-d.sub.6) .delta.
10.18 (broad s, 1H), 10.04 (broad s, 1H), 9.91 (broad s, 1H), 9.85
(broad s, 1H), 8.78 (d, J=1.2 Hz, 1H), 8.45 (d, J=5.7 Hz, 1H), 8.15
(s, 1H), 8.06 to 8.02 (m, 2H), 7.94 (d, J=2.7 Hz, 1H), 7.64 (d,
J=8.7 Hz, 2H), 7.54 to 7.52 (m, 3H), 7.20 (d, J=8.7 Hz, 2H), 7.12
(dd, J=5.7, 3.0 Hz, 1H); LC-MS m/z 443 [M+H].sup.+, RT=3.19
min.
Example 14
c-raf (raf-1) Biochemical Assay
The c-raf biochemical assay was performed with a c-raf enzyme that
was activated (phosphorylated) by Lck kinase. Lck-activated c-raf
(Lck/c-raf) was produced in Sf9 insect cells by co-infecting cells
with baculoviruses expressing, under the control of the polyhedrin
promoter, GST-c-raf (from amino acid 302 to amino acid 648) and Lck
(full-length). Both baculoviruses were used at the multiplicity of
infection of 2.5 and the cells were harvested 48 h post
infection.
MEK-1 protein was produced in Sf9 insect cells by infecting cells
with the baculovirus expressing GST-MEK-1 (full-length) fusion
protein at the multiplicity of infection of 5 and harvesting the
cells 48 hours post infection. Similar purification procedure was
used for GST-c-raf 302-648 and GST-MEK-1.
Transfected cells were suspended at 100 mg of wet cell biomass per
mL in a buffer containing 10 mM sodium phosphate, 140 mM sodium
chloride pH 7.3, 0.5% Triton X-100 and the protease inhibitor
cocktail. The cells were disrupted with Polytron homogenizer and
centrifuged 30,000 g for 30 minutes. The 30,000 g supernatant was
applied onto GSH-Sepharose. The resin was washed with a buffer
containing 50 mM Tris, pH 8.0, 150 mM NaCl and 0.01% Triton X-100.
The GST-tagged proteins were eluted with a solution containing 100
mM Glutathione, 50 mM Tris, pH 8.0, 150 mM NaCl and 0.01% Triton
X-100. The purified proteins were dialyzed into a buffer containing
20 mM Tris, pH 7.5, 150 mM NaCl and 20% Glycerol.
Test compounds were serially diluted in DMSO using three-fold
dilutions to stock concentrations ranging typically from 50 .mu.M
to 20 nM (final concentrations in the assay range from 1 .mu.M to
0.4 nM). The c-Raf biochemical assay was performed as a radioactive
filtermat assay in 96-well Costar polypropylene plates (costar
3365). The plates were loaded with 75 .mu.L solution containing 50
mM HEPES pH 7.5, 70 mM NaCl, 80 ng of Lck/c-raf and 1 .mu.g MEK-1.
Subsequently, 2 .mu.L of the serially diluted individual compounds
were added to the reaction, prior to the addition of ATP. The
reaction was initiated with 25 .mu.L ATP solution containing 5
.mu.M ATP and 0.3 .mu.Ci [33P]-ATP. The plates were sealed and
incubated at 32.degree. C. for 1 h. The reaction was quenched with
the addition of 50 .mu.L of 4% Phosphoric Acid and harvested onto
P30 filtermats (PerkinElmer) using a Wallac Tomtec Harvester.
Filtermats were washed with 1% Phosphoric Acid first and deionized
H.sub.2O second. The filters were dried in a microwave, soaked in
scintillation fluid and read in a Wallac 1205 Betaplate Counter
(Wallac Inc., Atlanta, Ga., U.S.A.). The results were expressed as
percent inhibition. % Inhibition=[100-(T.sub.ib(T.sub.i)].times.100
where T.sub.ib=(counts per minute with inhibitor)-(background)
T.sub.i=(counts per minute without inhibitor)-(background)
Example 15
flk-1 (Murine VEGFR-2) Biochemical Assay
This assay was performed in 96-well opaque plates (Costar 3915) in
the TR-FRET format. Reaction conditions are as follows: 10 .mu.M
ATP, 25 nM poly GT-biotin, 2 nM Eu-labelled phospho-Tyr Ab, 10 nM
APC, 7 nM flk-1 (kinase domain), 1% DMSO, 50 mM HEPES pH 7.5, 10 mM
MgCl.sub.2, 0.1 mM EDTA, 0.015% BRIJ, 0.1 mg/mL BSA, 0.1%
mercapto-ethanol). Reaction is initiated upon addition Of enzyme.
Final reaction volume in each well is 100 .mu.L. Plates are read at
both 615 and 665 nM on a Perkin Elmer Victor V Multilabel counter
at about 1.5-2.0 hours after reaction initiation. Signal is
calculated as a ratio: (665 nm/615 nm)*10000 for each well.
For IC.sub.50 generation against flk-1 kinase, test compounds were
added prior to the enzyme initiation. A 50-fold stock plate was
made with compounds serially diluted 1:3 in a 50% DMSO/50% dH2O
solution. A 2 .mu.L addition of the stock to the assay gave final
compound concentrations ranging from 10 .mu.M-4.56 nM in 1% DMSO.
The data were expressed as percent inhibition: %
inhibition=100-((Signal with inhibitor-background)/(Signal without
inhibitor-background))*100.
Compounds of examples 1-13 showed significant inhibition
(IC.sub.50<10 .mu.M) in either or both the c-raf and flk-1
biochemical assays.
* * * * *